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Ink set for inket printing |
An inkjet printing ink set giving good discharge stability is provided, which is able to deliver prints with a wide color reproduction gamut, especially pertaining to the yellow-red-magenta hue range, and with extremely low white ground tainting. The inkjet printing ink set of the present invention is for use in inkjet printing onto fabric made up of polyamide fibers, and is equipped with dye inks for at least the two colors yellow and magenta. Its special feature is that the dye ink for yellow contains 5 to 8% by weight of C.I. Acid Yellow 110 as its colorant and the dye ink for magenta contains 2.5 to 5% by weight of C.I. Acid Red 289 as its colorant. |
1. An ink set for use in inkjet printing onto fabric made up of polyamide fibers, which is equipped with dye inks for at least the two colors of yellow and magenta, wherein the dye ink for yellow contains 4 to 8% by weight of C. I. Acid Yellow 110 as its colorant, and the dye ink for magenta contains 2 to 5% by weight of C. I. Acid Red 289 as its colorant. 2. The inkjet printing ink set as in claim 1, which is additionally equipped with dye inks for orange and/or red. 3. The inkjet printing ink set as in claim 2, wherein the dye ink for orange contains 3 to 5% by weight of C. I. Acid Orange 56 as its colorant and the dye ink for red contains 5.5 to 8% by weight of C. I. Acid Red 249 as its colorant. 4. The inkjet printing ink set as in claim 1, which is additionally equipped with dye inks for black and/or cyan. 5. The inkjet printing ink set as in claim 4, wherein the dye ink for black contains 4 to 8% by weight of C.I. Acid Black 52 as its colorant and the dye ink for cyan contains 4 to 8% by weight of C. I. Direct Blue 87 as its colorant. 6. The inkjet printing ink set as claim 1, wherein each of the dye inks contains humectant, a water-soluble organic solvent (penetrant), a surfactant, and water. 7. The inkjet printing ink set as in claim 6, wherein the humectant is contained in the amount 4 to 40% by weight, the water-soluble organic solvent in the amount 0.5 to 15% by weight, and the surfactant in the amount 0.1 to 5% by weight, in the dye inks. 8. The inkjet printing ink set as in claim 6, wherein the surfactant is an acetylene glycol compound. 9. A printing method whereby the inkjet printing ink set as in claim 1 is used to perform inkjet printing onto fabric. 10. Prints produced using the printing method as in claim 9. 11. The inkjet printing ink set as in claim 2, which is additionally equipped with dye inks for black and/or cyan. 12. The inkjet printing ink set as in claim 3, which is additionally equipped with dye inks for black and/or cyan. 13. The inkjet printing ink set as claim 2, wherein each of the dye inks contains humectant, a water-soluble organic solvent (penetrant), a surfactant, and water. 14. The inkjet printing ink set as claim 3, wherein each of the dye inks contains humectant, a water-soluble organic solvent (penetrant), a surfactant, and water. 15. The inkjet printing ink set as claim 4, wherein each of the dye inks contains humectant, a water-soluble organic solvent (penetrant), a surfactant, and water. 16. The inkjet printing ink set as claim 5, wherein each of the dye inks contains humectant, a water-soluble organic solvent (penetrant), a surfactant, and water. 17. The inkjet printing ink set as in claim 7, wherein the surfactant is an acetylene glycol compound. 18. A printing method whereby the inkjet printing ink set as in claim 2, is used to perform inkjet printing onto fabric. 19. A printing method whereby the inkjet printing ink set as in claim 3, is used to perform inkjet printing onto fabric. 20. A printing method whereby the inkjet printing ink set as in claim 4, is used to perform inkjet printing onto fabric. |
<SOH> BACKGROUND ART <EOH>The techniques generally used for printing text and/or images onto woven, non-woven or other types of fabric made from various kinds of fibers are the screen printing method, the roller printing method and certain others. However such printing methods entail the bothersome work of preparing a trace or plate for each design, because of which they are not suited to realize low-cost processes, and are unsuited to the production of numerous different items in small volumes. To eliminate such drawbacks of these traditional printing methods, a method (ink jet printing) has been advanced and developed for practical use, whereby designs are read into a scanner or other image input device, undergo image processing by a computer, and are printed onto the fabric according to the image data by means of the ink jet reproduction technique. This ink jet reproduction technique performs printing by projecting droplets of ink in a jet and causing them to adhere to the substrate. It is a technique that has come to be widely employed in printing fields where the substrate is paper or similar, due to merits including the fact that it readily provides high-quality full-color images. Ink jet printing generally employs an ink set equipped with dye inks for the four colors: black, yellow, magenta and cyan, and there is a desire for the development of ink sets that would provide prints with high image quality and color fastness while also achieving excellent discharge stability. To date, various inkjet printing ink sets have been put forward in response to this desire, but have had the problem that they are unable to fully reproduce the range of hues that is obtainable with traditional printing methods such as the screen process. Examples of ink sets intended to resolve this problem are seen in Japan Laid-Open Patent Applications H6-25576, H6-57654 and H7-3666, which put forward ink sets composed of multiple dye inks yielding particular ranges for each of chromaticness indexes a* and b* defined in an L*a*b* color space (CIE 1976) on the fabric. However, the ink sets described in these laid-open patent applications have poor color reproduction in the yellow-red-magenta hue range, and an inkjet printing ink set with a color reproduction gamut of adequate range has yet to be provided. A further example of a response to the aforementioned desire is the ink set put forward in Japan Laid-Open Patent Application H8-259832, where each of the inks for the colors of yellow, magenta and cyan contains a particular acidic dye. However, prints produced using ink jets of this kind suffer from a phenomenon that is known as “color bleed,” the intermixing of different colors at their boundaries, and thus have not been satisfactory in regard to image quality. Moreover, fabrics printed by inkjet printing are generally subjected to a process (“soaping process”) whereby unfixed dye is washed off with a hot soap solution or similar, during which the washed-off dye is liable to adhere to and taint the white portions of the printed fabric, resulting in “white ground tainting.” Accordingly, an object of the present invention is to provide an inkjet printing ink set that is able to deliver a broad color reproduction gamut—particularly for the yellow-red-magenta hue range—and prints with extremely low white ground tainting, together with good discharge stability. Another object of the present invention is to provide an inkjet printing ink set that is able to deliver prints with extremely low color bleed and white ground tainting, together with good discharge stability. |
Super absorbent driven dispenser |
A device (1) for causing the displacement of a piston (36) or diaphragm (4) comprising an enclosure containing unsaturated super-absorbent material (38) and including a piston or diaphragm; and a fluid inlet into the enclosure adapted to allow fluid into the enclosure and restrain the super-absorbent material form escaping from the enclosure; such that as fluid is allowed into the enclosure, it is absorbed by the super-absorbent material which swells against the piston (36) or diaphragm (4), causing displacement thereof. The piston (36) or diaphragm (4) may form at least one wall of the enclosure. Alternatively diaphragm may be a membrane of elastomeric material within the enclosure. The invention may be used in a drug delivery system, a air freshener, or a lavatory cleaner. |
1. A device for causing the displacement of a piston or diaphragm comprising: an enclosure containing unsaturated super-absorbent material and including a piston or diaphragm; and a fluid inlet into the enclosure adapted to allow fluid into the enclosure and restrain the super-absorbent material form escaping from the enclosure; the arrangement being such that as fluid is allowed into the enclosure, it is absorbed by the super-absorbent material which swells against the piston or diaphragm, causing displacement thereof. 2. A device for causing the displacement of a piston or diaphragm as claimed in claim 1, wherein the piston or diaphragm forms at least one wall of the enclosure. 3. A device for causing the displacement of a piston or diaphragm as claimed in claim 1, wherein the piston or diaphragm is a membrane of elastomeric material within the enclosure, dividing the enclosure into two sections, one for holding the unsaturated super-absorbent material, and the other for holding liquid or the like to be displaced. 4. A device for causing the displacement of a diaphragm as claimed in claim 1, wherein the diaphragm is a bag within the enclosure, sealed at or near an outlet of the enclosure, the bag being fillable with a liquid to be displaced. 5. A device for causing the displacement of a piston or diaphragm as claimed in any preceding claim, wherein the volume of water is controlled to determine the expansion of the super-absorbent material. 6. A device for causing the displacement of a piston or diaphragm as claimed in preceding claim, wherein the rate of expansion of the super-absorbent material is controlled by controlling the purity of the water. 7. A device for causing the displacement of a piston or diaphragm as claimed in any preceding claim, wherein the super-absorbent material is in granular form. 8. A device for causing the displacement of a piston or diaphragm as claimed in any one of claims 1 to 6, wherein the super-absorbent material is in fibrous form. 9. A device for causing the displacement of a piston or diaphragm as claimed in any preceding claim, wherein the enclosure is an extension of a cap into a body, the body being adapted to hold fluid, and the fluid inlet being between the extension and a base of the body on lifting of the extension. 10. A device for causing the displacement of a piston or diaphragm substantially as hereinbefore described with reference to FIGS. 1 to 3, FIGS. 4 and 5, FIGS. 6 and 7 and FIG. 8 of the accompanying drawings. |
Configurations and method for improved gas removal |
A plant includes an absorber (103) that operates in a gas phase supercritical region and removes an acid gas from a feed stream (9) at high recovery of the feed stream (10) while producing a high purity acid gas stream (36). Particularly preferred plants include gas purification plants that receive a feed gas with at least 5 mol % carbon dioxide at a pressure of at least 3000 psi. |
1. A plant comprising an absorber that operates in a gas phase supercritical region and receiving a dehydrated feed gas comprising at least 5 mol % carbon dioxide, wherein a physical solvent absorbs at least part of the carbon dioxide in the absorber to form a carbon dioxide stream containing at least 95 mol % carbon dioxide. 2. The plant of claim 1 wherein the feed gas is cooled to a temperature above a hydrate point of the feed gas and wherein the cooled feed gas is dehydrated in a dehydration unit before entering the absorber. 3. The plant of claim 1 wherein the feed gas has a pressure of at least 3000 psi. 4. The plant of claim 1 wherein the feed gas comprises natural gas. 5. The plant of claim 1 wherein the physical solvent comprises propylene carbonate. 6. The plant of claim 1 wherein the absorber forms a rich solvent that is depressurized, thereby providing cooling of the carbon dioxide stream and further providing a first hydrocarbon recycle stream that is fed into the absorber. 7. The plant of claim 6 wherein the depressurized rich solvent is further depressurized, thereby providing cooling of the solvent in the absorber and further providing a second hydrocarbon recycle stream that is fed into the absorber. 8. The plant of claim 7 wherein the further depressurized rich solvent is let down and separated in a separator to form a lean solvent and the carbon dioxide stream. 9. The plant of claim 7 wherein a compressor compresses the first and second hydrocarbon recycle stream and wherein the compressed first and second hydrocarbon recycle streams are combined with the dehydrated feed gas. 10. The plant of claim 8 wherein the lean solvent is treated in a vacuum separator using lean gas thereby regenerating the physical solvent for the absorber and further generating a fuel gas. 11. The plant of claim 2 wherein the absorber produces a product gas, and wherein the feed gas is cooled by the product gas. 12. The plant of claim 11 wherein the feed gas comprises natural gas, and wherein the product gas comprises at least 99% of the natural gas. 13. The plant of claim 1 wherein the carbon dioxide stream is employed in an enhanced oil recovery. 14. A plant comprising: an absorber that receives a feed gas at a pressure of at least 1000 psi and comprising at least 5 mol % carbon dioxide, wherein the absorber employs a physical solvent to produce a rich solvent and a product gas that is at least partially depleted from the carbon dioxide; a first turbine that depressurizes the rich solvent and a first separator that separates a first hydrocarbon portion from the depressurized rich solvent, thereby producing a first hydrocarbon recycle stream and a first rich solvent; a second turbine that further depressurizes the first rich solvent and a cooler that employs the further depressurized first solvent to cool the physical solvent to maintain a bottom temperature of the absorber in a desired temperature range; and wherein the further depressurized first solvent is separated in a second separator that separates a second hydrocarbon portion from the further depressurized first solvent, thereby producing a second hydrocarbon recycle stream and a second rich solvent, and wherein the first and second hydrocarbon recycle streams are fed into the absorber. 15. The plant of claim 14 wherein the feed gas is at least partially dehydrated. 16. The plant of claim 15 wherein he feed gas is cooled by the product gas and dehydrated in a dehydration unit before entering the absorber. 17. The plant of claim 14 wherein the first rich solvent is employed as a refrigerant to cool a carbon dioxide stream. 18. The plant of claim 14 wherein the second rich solvent is further depressurized to remove at least part of the carbon dioxide, thereby forming a lean solvent. 19. The plant of claim 18 wherein the lean solvent is further processed in a vacuum separator that employs a lean gas and produces a fuel gas, thereby regenerating the physical solvent. 20. The plant of claim 18 wherein the physical solvent comprises propylene carbonate. 21. The plant of claim 18 wherein the carbon dioxide is employed in an enhanced oil recovery. 22. The plan of claim 14 further comprising a stripper having at least two stripping sections separated by a collecting tray, and wherein the stripper regenerates the physical solvent 23. A method of operating a plant, comprising; providing an absorber that receives a feed gas at a pressure of at least 1000 psi wherein the feed gas comprises an acid gas; removing at least pan of the acid gas from the feed gas using a physical solvent, thereby forming a product gas and a rich solvent; reducing pressure of the rich solvent to form a hydrocarbon recycle stream that is fed into the absorber, thereby producing a depressurized rich solvent; using the depressurized rich solvent to cool the physical solvent, thereby maintaining a bottom temperature of the absorber in a desired temperature range and forming a heated depressurized rich solvent; and separating the heated depressurized rich solvent into a stream containing at least a portion of the acid gas, thereby forming a lean solvent. 24. The method of claim 23 wherein the fed gas has a pressure of between about 3000 psi and about 5000 psi. 25. The method of claim 23 wherein the acid gas is carbon dioxide. 26. The method of claim 23 wherein the physical solvent comprises propylene carbonate. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Acid gas removal from various gas streams, and especially removal of carbon dioxide from natural gas streams has become an increasingly important process as the acid gas content of various gas streams increases. For example, the carbon dioxide concentration in natural gas from enhanced oil recovery will typically increase from 10% to about 60%. There are numerous processes for acid gas removal known in the art, and all or almost all of those may be categorized into one of three categories. In the first category, one or more membranes are used to physically separate the acid gas from a gaseous feed stream. A typical membrane system includes a pre-treatment skid and a series of membrane modules. Membrane systems are often highly adaptable to accommodate treatment of various gas volumes and product-gas specifications. Moreover, membrane systems are relatively compact, therefore rendering membrane systems an especially viable option for offshore gas treatment. However, membrane systems are susceptible to deterioration from heavy hydrocarbons content in the feed gas. Moreover, carbon dioxide removal to relatively low carbon dioxide content typically requires multiple stages of membrane separators and recompression between stages, which tend to be relatively expensive. In the second category, a chemical solvent is employed that reacts with the acid gas to form a (typically non-covalent) complex with the acid gas. In processes involving a chemical reaction between the acid gas and the solvent, the crude gases are typically scrubbed with an alkaline salt solution of a weak inorganic acid as, for example, described in U.S. Pat. No. 3,563,695, or with alkaline solutions of organic acids or bases as, for example, described in U.S. Pat. No. 2,177,068. Such chemical reaction processes generally require heat regeneration and cooling of the chemical solvents, and often involve recirculation of large amounts of chemical solvent. Moreover, the quantity of chemical solvent required to absorb increasing amounts of acid gases generally increases significantly, thus making use of chemical solvents problematic where the acid gas content increases over time in the feed gas. In the third category, a physical solvent is employed for removal of acid gas from a feed gas, wherein the acid gas does react in an appreciable amount with the solvent. The physical absorption of the acid gas predominantly depends upon use of solvents having selective solubilities for the particular acid gas (e.g., CO 2 or H 2 S) gaseous components to be removed and is further dependent upon pressure and temperature of the solvent. For example, methanol may be employed as a low-boiling organic physical solvent, as exemplified in U.S. Pat. No. 2,863,527. However, the energy input requirements for cooling are relatively high, and the process generally exhibits greater than desired methane and ethane absorption, thereby necessitating large energy inputs for recompression and recovery. Alternatively, physical solvents may be operated at ambient or slightly below ambient temperatures, including propylene carbonates as described in U.S. Pat. No. 2,926,751 and those using N-methylpyrrolidone or glycol ethers as described in U.S. Pat. No. 3,505,784. While such solvents may advantageously reduce cooling requirements, most propylene carbonate-based absorption processes are limited to absorption pressures of less than 1000 psi (i.e., at sub-critical pressure). In further known methods, physical solvents may also include ethers of polyglycols, and specifically dimethoxytetraethylene glycol as shown in U.S. Pat. No. 2,649,166, or N-substituted morpholine as described in U.S. Pat. No. 3,773,896. While use of physical solvents avoids at least some of the problems associated with chemical solvents and/membranes, various new difficulties arise. Among other things, carbon dioxide removal from high pressure feed gases is often limited to sub-critical pressures. Furthermore, as the water content increases, freezing may occur in the solvent circuit, thus necessitating a relatively high temperature and thereby reducing the efficiency of the absorption process. In another aspect, physical solvent generally requires steam or external heat for solvent regeneration in order to produce a very lean solvent suitable for removal of acid gas to the ppm level. Thus, although various configurations and methods are known to remove acid gases from a feed gas, all or almost all of them suffer from one or more disadvantages. Therefore, there is still a need to provide methods and configurations for improved acid gas removal. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is directed towards methods and configurations of a plant in which an acid gas component is removed from a feed stream, wherein the feed stream comprises at least 5 mol % carbon dioxide and has a pressure of at least 1000 psi. In one aspect of the inventive subject matter, a plant comprises an absorber operating in a gas phase supercritical region and receiving a dehydrated feed gas comprising at least 5 mol % carbon dioxide, wherein a physical solvent absorbs at least part of the carbon dioxide in the absorber to form a carbon dioxide stream containing at least 95 mol % carbon dioxide. The feed gas in such configurations is preferably cooled to a temperature above the hydrate point of the feed gas for removal of the majority of the water content, and the cooled feed gas is then dehydrated in a dehydration unit before entering the absorber. It is further preferred that the feed gas (e.g., natural gas) has a pressure of between about 3000 psig to about 5000 psig. Particularly preferred physical solvents include propylene carbonate, and it is contemplated that the absorber forms a rich solvent from the physical solvent that is depressurized, thereby providing cooling of a carbon dioxide stream and further providing a first hydrocarbon recycle stream that is fed into the absorber. In yet other aspects, the depressurized rich solvent is further depressurized, thereby providing cooling of the solvent in the absorber and further providing a second hydrocarbon recycle stream that is fed into the absorber. The further depressurized rich solvent may then be let down and can be separated in a separator to form a lean solvent and the carbon dioxide stream. Where hydrocarbon recycle streams are generated, it is contemplated that such streams will be compressed in a compressor to the pressure of the absorber, and that the compressed combined recycle stream can then be mixed with the dehydrated feed gas. The lean solvent in preferred configurations may be treated in a vacuum stripping unit using a stripping gas, thereby regenerating the physical solvent for the absorber and further generating a fuel gas, and the product gas from the absorber may cool the feed gas. While not limiting to the inventive subject matter, it is further preferred that the product gas comprises at least 99% of the natural gas, and that the carbon dioxide stream is employed in an enhanced oil recovery. Where a very low acid gas specification is required of the product gas, it is contemplated that the lean solvent in preferred configurations comprises at least two streams from the vacuum stripper. A partially stripped solvent is generated in the upper section of the stripping column that is pumped to the middle section of the absorber, and an ultra stripped lean solvent from the lower section of the stripping column is pumped and cooled and sent to the top of the absorber. Such ultra stripped lean solvent configurations produce a product gas that will meet a ppm level acid gas specification that is previously achievable only with heat regenerated solvent. Consequently, in another aspect of the inventive subject matter, a gas treating plant may comprise an absorber receiving a feed gas at a pressure of at least 1000 psi and comprising at least 5 mol % carbon dioxide, wherein the absorber employs a physical solvent to produce a rich solvent and a product gas that is at least partially depleted from the carbon dioxide. Suitable plants may further include a first turbine that depressurizes the rich solvent and a first separator that separates a first hydrocarbon portion from the depressurized rich solvent, thereby producing a first hydrocarbon recycle stream and a first rich solvent, and a second turbine that further depressurizes the first rich solvent and a cooler that employs the further depressurized first solvent to cool the physical solvent to maintain a bottom temperature of the absorber in a desired temperature range. In such configurations, it is preferred that the further depressurized first solvent is separated in a second separator that separates a second hydrocarbon portion from the further depressurized first solvent, thereby producing a second hydrocarbon recycle stream and a second rich solvent, and wherein the first and second hydrocarbon recycle streams are fed into the absorber. Thus, a method of operating a plant may include one step in which an absorber is provided that receives a feed gas at a pressure of at least 1000 psi and comprises an acid gas. In another step, at least part of the acid gas is removed from the feed gas using a physical solvent, thereby forming a product gas and a rich solvent, and in still another step, the pressure of the rich solvent is reduced to form a hydrocarbon recycle stream that is fed into the absorber, thereby producing a depressurized rich solvent. In a further step, the depressurized rich solvent is used to cool the physical solvent, thereby maintaining a bottom temperature of the absorber in a desired temperature range and forming a heated depressurized rich solvent, and in yet another step, the heated depressurized rich solvent is separated into a stream containing at least a portion of the acid gas, thereby forming a lean solvent. Yet in another step, the lean solvent is depressurized to a vacuum stripper, which produces a partially stripped lean solvent and an ultra stripped lean solvent that are fed to two different locations in the absorber. The stripper overhead gas can be used as a plant fuel gas. |
Lectin-like domain of thrombomodulin and its therapeutic use |
In the present invention the in vivo role of the N-terminal lectin-like domain of thrombomodulin was studied by using homologous recombination in murine ES cells to create mutant mice that lack this region of thrombomodulin. Phenotypic analysis shows that said mice respond identically to their wild type littermates following pro-coagulant challenges meaning that the protein C pathway is not altered by the mutation. However, following several inflammatory stimuli, it was observed that the mutant mice showed an elevated neutrophil extravasation in several organs. In the present invention it is found that leukocyte adhesion could be abrogated by addition of recombinant lectin-domain meaning that said domain has direct anti-inflammatory properties which means that the lectin-like domain can be used to manufacture a medicament useful for the treatment of a variety of inflammatory disease processes. |
1. A polypeptide consisting essentially of an amino acid sequence corresponding to SEQ ID NO: 1 or fragments or homologues thereof for use as a medicament. 2. A fragment according to claim 1 as depicted in SEQ ID NO:2, 3, 4, 5, 6, 7, or 8. 3. A method of using a polypeptide according to claims 1 for the manufacture of a medicament to prevent and/or to treat inflammation. 4. The method of using a polypeptide according to claim 3 wherein said inflammation is a result of ischemia-reperfusion injury. 5. The method of using a polypeptide according to claim 3 to prevent leukocyte adhesion and/or invasion. 6. The method of using a polypeptide according to claim 5 wherein said leukocyte is a neutrophil. 7. A polypeptide or a fragment thereof according to claims 1 2 which is a recombinant peptide. 8. A recombinant DNA vector having an insert consisting of a DNA sequence encoding the recombinant polypeptide or a fragment thereof according to claims 9. A host cell comprising the recombinant DNA vector according to claim 8. 10. A process for producing a polypeptide or a fragment thereof according to claim 7 which comprises the following steps: preparing a DNA fragment comprising a nucleotide sequence which encodes said polypeptide, incorporating said DNA fragment into a recombinant DNA vector which contains said DNA fragment and is capable of undergoing replication, transforming a host cell with said recombinant DNA fragment to isolate a transformant which can express said polypeptide, and culturing said transformant to allow the transformant to produce said polypeptide and recovering said polypeptide from resulting cultured mixture. 11. A method of using a polypeptide according to claim 2 for the manufacture of a medicament to prevent and/or to treat inflammation. 12. The method of using a polypeptide according to claim 11 wherein said inflammation is a result of ischemia-reperfusion injury. 13. The method of using a polypeptide according to claim 11 to prevent leukocyte adhesion and/or invasion. 14. The method of using a polypeptide according to claim 13 wherein said leukocyte is a neutrophil. 15. A polypeptide or a fragment thereof according to claim 2 which is a recombinant peptide. 16. A recombinant DNA vector having an insert consisting of a DNA sequence encoding the recombinant polypeptide or a fragment thereof according to claim 2. 17. A host cell comprising the recombinant DNA vector according to claim 16. 18. A process for producing a polypeptide or a fragment thereof according to claim 15 which comprises the following steps: preparing a DNA fragment comprising a nucleotide sequence which encodes said polypeptide, incorporating said DNA fragment into a recombinant DNA vector which contains said DNA fragment and is capable of undergoing replication, transforming a host cell with said recombinant DNA fragment to isolate a transformant which can express said polypeptide, and culturing said transformant to allow the transformant to produce said polypeptide and recovering said polypeptide from resulting cultured mixture. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Although it has long been recognized that the coagulation system plays a role in modulating inflammation, it is only recently that the impact of this contribution has been appreciated and that some of the molecular links have been established. In this respect, the protein C anticoagulant pathway is particularly relevant. In addition to its well-characterized role in modulating thrombin generation, this system, composed of a complex of soluble and membrane-associated proteins, plays an integral part in regulating the response to selected inflammatory agents (reviewed in 44 ). Substantial clinical data have revealed that patients with severe sepsis have significantly diminished levels of protein C and protein S, and the extent of suppression of protein C may correlate with clinical outcome 49 . Activated protein C (APC) appears to modulate the inflammatory response by several mechanisms, including inhibiting polymorphonuclear cell (PMN) activation and elastase release, blocking PMN interactions with selecting, and preventing cytokine release by monocytes 48,52-55 . More recently, the endothelial cell protein receptor (EPCR), a cofactor that enhances activation of protein C by thrombin-thrombomodulin, has also been found to modulate the function of APC in inflammation. Moreover, inhibition of the interaction of APC/PC with EPCR in vivo resulted in an increased inflammatory response following E. coli infusions in baboons 56 . Further links between EPCR and inflammation, although not yet fully delineated, are being explored as Esmon and coworkers have reported that a soluble form of EPCR is released during sepsis 57 , interferes with activation of protein C, and binds to a receptor on activated neutrophils that is the autoantigen in Wegener's granulomatosis 58,59 . Another particularly relevant player in the anticoagulant system is thrombomodulin (TM), a critical cofactor in the activation of protein C, and a widely expressed glycoprotein receptor for thrombin. With the cloning and sequencing of the gene for thrombomodulin 1 , the putative structural organization of the protein and the regions responsible for its anticoagulant and anti-fibrinolytic function have been elucidated. Mature single-chain TM in the human is 557 amino acids long and is structurally divided into five domains. The N-terminal region (residues 1-226) 2 has a module (residues 1-154) with homology to the lectin domains of the hepatic asialoglycoprotein receptor and IgE, as well as to members of the selectin family. Although controversial, in vitro analyses suggest that this domain is required for constitutive internalization of the receptor in some cells 5,6 . From residues 155 to 226, there is a hydrophobic region which may be associated with the plasma membrane and which contains two potential sites for O-linked glycosylation. The next domain is comprised of six epidermal growth factor (EGF)-like repeats, the last 3 or 4 of which are necessary for activation of TAFI or protein C, respectively, by thrombin. The function of the other EGF-like repeats is unknown. The third domain between the EGF-like repeats and the membrane-spanning region is serine/threonine rich and contains four potential sites for O-linked glycosylation, to one of which is attached a chondroitin sulfate, important for full anticoagulant activity of TM. Fourthly, there is a highly conserved transmembrane domain, and fifthly a short cytoplasmic tail that contains potential sites of phosphorylation, and a single cysteine that may mediate multimerization of the molecule. It has been shown that TM is important in regulating the inflammatory process via the anticoagulant pathway. The downregulation of vascular endothelial cell TM by inflammatory cytokines—an effect mirrored by the expression of cellular EPCR—directly impairs the generation of APC. The protein C co-factor function of TM is also impaired in the face of inflammation, as activated PMNs release lysosomal proteases and oxidants that result in proteolysis of the receptor and oxidation of a critical methionine within the EGF-like repeats of TM that inactivates the function of glycoprotein for protein C activation. Several additional lines of evidence support a role for TM as an anti-inflammatory agent. Recombinant soluble forms of TM, most of which were composed of the entire extramembranous regions, were used to prevent endotoxin-induced pulmonary accumulation of leukocytes and ARDS, organ failure, or lethality in small animal models 54,63,64 . Adenovirus-mediated gene transfer of TM in a rabbit restenosis model was not only effective in reducing restenosis, but also resulted in decreased inflammation and extravasation of leukocytes 22 . In a spinal cord compression-induced injury model in rats, recombinant soluble TM provided neuroprotection, with reduction in leukocyte accumulation and cytokine mRNA expression 65 . In each of these studies, the improved outcomes following TM administration were attributed to enhanced activation of protein C, while the possibility that other domains of TM might contribute to the apparent anti-inflammatory effect was never considered. In the present invention we have determined the in vivo function of the N-terminal lectin-like domain of TM by generating mice lacking this domain and we have shown that addition of the recombinant N-terminal lectin-like domain provides the vascular endothelium with natural anti-inflammatory properties by interfering with leukocyte adhesion. (1) TM is a known molecule, (2) the EGF-regions of TM are known to have anti-coagulant (and indirectly anti-inflammatory) activity. However, the current invention surprisingly demonstrates that the lectin-like region of TM has an anti-inflammatory function. Indeed, since it has been shown in the art that several members of the C-type lectin family (to which the lectin-like domain of TM belongs) function to enhance leukocyte adhesion one would expect that the lectin-like domain of TM has rather a pro-inflammatory function. |
Lectin protein prepared from maackia fauriei, process for preparing the same and the use thereof |
The present invention relates to a lectin protein, designated MFA isolated and purified from the bark of the Korean legume Maackia fauriei, process for preparing the same and the use thereof. This protein can be used as reagents in the study of carbohydrate binding proteins as well as to examine the distribution of N-acetylneuraminic acid in cancer cells owing to its capability that specifically recognizes N-acetylneuraminic acid which plays important structural and functional roles in the expressions of various cells or oligosaccharide terminal residue of glycoconjugates, and, in addition, used as an anti-cancer drug in view of its anti-proliferation effect against various cancers such as breast cancer, melanoma, hepatoma, etc. |
1. A lectin MFA (Maackia fauriei agglutinin) protein extracted from the Korean legume Maackia fauriei characterized in that it has a molecular weight of approximately 30.0 kDa by electrophoresis and mass spectrometry and its N-terminal amino acid sequence is represented in sequence no. 1 of the sequence listing. 2. The MFA protein as claimed in claim 1, wherein it specifically binds to N-acetylneuraminic acid or a glycoprotein including N-acetylneuraminic acid by hemagglutination reaction, and hemagglutination-inhibition reaction. 3. A process for preparing a lectin MFA protein from the bark of the Korean plant Maackia fauriei , which comprising the steps of: removing the outer coat from the bark of Maackia fauriei , homogenizing the bark and mixing the homogenate with an aqueous NaCl solution under stirring, filtering the precipitate with a filter, applying the filtrate onto column chromatography to give the crude fractions and further purifying the active fraction through a fetuin-affinity column to give the lectin MFA protein. 4. A diagnosing agent for a disease in which the structure of carbohydrates within the cells contains N-acetylneuraminic acid and its distribution is changed due to invasion of diseases, which comprises the lectin MFA according to claim 1. 5. A drug conjugate in which the lectin MFA protein according to claim 1 as a carrier protein for local transport of a drug and a selective drug as an active agent are bonded, 6. A use of the lectin MFA according to claim 1 in the research material of immunobiochemistry or in agglutination of cancer cells. 7. An anti-proliferation agent for cancer cells comprising the lectin MFA protein according to claim 1 as an active agent. 8. The anti-proliferation agent for cancer cells as claimed in claim 7, wherein the cancer includes breast cancer, melanoma and hepatoma. |
<SOH> BACKGROUND ART <EOH>Recently, as the techniques for elucidating and controlling the structure and function of a biological material have been rapidly developed in the genetic engineering or protein engineering, the understanding of vital phenomenon is progressed at the gene and protein levels, and at the same time, carbohydrates such as oligosaccharides in the living body are also considered as an essential subject in elucidating the regulatory mechanism of the living body. In order to advance the related researches, the studies on the effect of oligosaccharides on the structure and function of the living body and on the effect on intercellular communication, etc. shall be systemically carried out. Oligosaccharides, an essential component constituting the living body play essential roles in maintaining biological activities such as cellular adhesion, intercellular communication, and morphogenesis of individual tissue by forming a complex molecule such as glycoproteins or glycolipids conjugated with proteins or lipids. The functions of oligosaccharides known hitherto are mainly classified into three kinds: First, oligosaccharides bind to other proteins, thus playing an important role in specific recognition of cells by the interaction. The next is the case where oligosaccharide bound to protein itself significantly modifies the inherent function of the protein. The example thereof includes N-CAM which is the cellular adhesive molecule of oligosaccharide specifically expressing in the brain such as polysialic acid. Third, most of proteins are glycoproteins in which oligosaccharide is bound to the protein via Asn or Ser/Thr of the protein itself by which activate functions of the protein. That is, oligosaccharide is considered to be an important material which results in change of function by itself or by the combination with other biological materials. Intercellular recognition occurs by way of carbohydrates on the cell membrane surface, and in order to mediate this process molecules specifically recognizing the carbohydrates should exist on the cell membrane surface. For example, a carbohydrate-binding protein such as lectin is necessary on the cell membrane surface. Korean legume Maackia fauriei , which belongs to the family Leguminosae is a deciduous forest tree and a specialty plant of Korea and is distributed in large amounts throughout the height 1,100-1800 m of the Cheju island in Korea. However, the biochemical researches thereon are very poor. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>Further objects and advantages of the invention can be more fully understood from the following detailed description taken in conjunction with the accompanying drawings in which: FIG. 1 is a graph showing a UV (280 nm) spectroscopic chromatogram and the hemagglutination response by a Sepharose CL-6B column of the extract containing the lectin MFA prepared by the present invention; FIG. 2 is a graph showing a UV (280 nm) spectroscopic chromatogram and hemagglutination response by a fetuin-affinity column of the lectin MFA prepared by the present invention; FIG. 3 is a photograph showing an electrophoretic analysis of the lectin MFA prepared by the present invention, in which lane 1 is a molecular weight marker (66, 45, 36, 29, 24, 20.1, 14.2 kDa); lane 2 is a Sepharose CL-6B eluate of Maackia fauriei (fraction nos. 9 to 17); lane 3 is the lectin MFA of Maackia faurie by a fetuin-affinity column(fraction nos. 53 to 58) and lane 4 is unbound eluate by the fetuin-affinity column(fraction nos. 8 to 16); FIG. 4 is a graph showing the results of a UV (280 nm) spectroscopic chromatogram by a gel filtration of MFA prepared by the process of the invention, in which graph (a) represents an eluate of the lectin MFA of Maackia faurie on a Superdex-200 HR column(fraction nos. 9 to 17), (b) represents an eluted lectin MFA of Maackia faurie on a fetuin-affinity column(fraction nos. 53 to 58) and graph (c) represents unbound eluate by the fetuin-affinity column(fraction nos. 8 to 16); FIG. 5 is a graph showing the molecular weight by the gel filtration column of the lectin MFA prepared by the process of the invention. Molecular weight marker(200, 150, 66, 29, 12.4 kDa); and FIG. 6 is a graph showing the molecular weight by MALDI-TOF mass spectrometer of the lectin MFA prepared by the process of the invention. detailed-description description="Detailed Description" end="lead"? |
Method for coloring cellulosic materials using cationic pigment dispersion |
A method of coloring a cellulosic material which includes a) dispersing pulped cellulosic material into water; and b) coloring the pulped cellulosic material by adding a cationic dispersion to the water, where the dispersion includes: (i) at least one pigment; (ii) water; and (iii) at least one quaternary salt of a styrene maleimide imide resin in an amount effective to disperse the pigment. The cationic dispersion may be prepared by (i) mixing, at 500 to 10,000 rpm, at least one pigment; water; and either (a) at least one a quaternary salt of a styrene maleimide imide resin or (b) at least one styrene maleimide imide resin in combination with at least one weak acid, thereby forming a dispersion premix; (ii) milling the dispersion premix in a mixer filled with ceramic, metal or glass beads for a period of time sufficient to reduce pigment agglomerates to primary particles, thereby forming a nonstandardized dispersion; and (iii) standardizing the dispersion against a color standard by adding water. The resulting cationic dispersion can be used to color cellulosic materials such as cotton and paper. |
1. A method of coloring a cellulosic material, comprising a) dispersing pulped cellulosic material into water; and b) coloring said pulped cellulosic material by adding a cationic dispersion into said water, wherein said dispersion comprises: (i) at least one pigment; (ii) at least one dispersing agent comprising a quaternary salt of a styrene maleimide imide resin; and (iii) water. 2. A cationic dispersion, comprising: (i) at least one pigment; (ii) at least one dispersing agent comprising a quaternary salt of a styrene maleimide imide resin; and (iii) water. 3. The cationic dispersion of claim 2, wherein said pigment is at least one organic pigment selected from the group consisting of phthalocyanine green, phthalocyanine blue, carbazole violet, toluidine red, Dalamar yellow, Watchung red and diketopyrrolopyrrole, quinacridone red, quinacridone yellow, quinacridone violet and arylide yellow. 4. The cationic dispersion of claim 2, wherein said organic pigment is a phthalocyanine. 5. The cationic dispersion of claim 2, wherein said pigment is at least one inorganic pigment selected from the group consisting of red oxide, yellow oxide, black iron oxide, cobalt blue, carbon black and bismuth vanadate. 6. The cationic dispersion of claim 2, further comprising at least one member of the group consisting of a surfactant, a biocide and a viscosity control agent. 7. The cationic dispersion of claim 2, wherein said pigment comprises primary particles. 8. A colored cellulosic material, consisting essentially of pigment particles coated with a styrene maleimide imide resin; said coated particles fixed on fibers of a cellulosic material. 9. The colored cellulosic material of claim 8, wherein said cellulosic material is selected from at least one member of the group consisting of paper and cotton. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention This invention relates to a method of coloring cellulosic materials such as paper pulp and cotton. More particularly, this invention relates to a method of coloring cellulosic materials using a cationic dispersion which contains at least one pigment, water, and at least one dispersing agent comprising a quaternary salt of a styrene maleimide imide resin. 2. Description of the Prior Art Papermaking is a well-known process in which a cellulosic material, typically obtained from wood, is mechanically or chemically pulped, dispersed in water, formed into a planar sheet, dried and wound onto a roll for later use. The paper may be sized to modify its surface characteristics, particularly water penetration, which is important for writing and printing grades of paper. Additives such as fillers and optical brighteners may be added to the pulp prior to sheet formation. Colorants such as dyes or pigments may also be added during the papermaking process, either by coloring the paper pulp, or applying the colorant to the paper surface by dip coating, spraying or pad printing. Pulp coloration is the most widely used type of paper coloration. “Substantivity” is the ability of a dye or pigment to be adsorbed by cellulose fibers from an aqueous medium. “Affinity” is the capability of a dye or pigment to be bound to cellulose fibers. Cellulosic materials are slightly anionic in water due to partly dissociated carboxylic acid and other functional groups. Some chemically treated pulps may also contain sulfonate groups. The anionic character of cellulosic materials in water affects the substantivity and affinity of dyes and pigments for paper. Thus, anionic dyes such as acid and anionic direct dyes will typically require the addition of fixing agents to overcome electrostatic repulsion from the anionic cellulose fibers. Cationic dyes such as basic and cationic direct dyes will be electrostatically attracted to the anionic cellulose fibers, but may still require fixing agents to achieve acceptable substantivity and affinity. Pigments have not enjoyed the field of coloring paper. about 60% of the paper market, and acid dyes and pigments make up the remainder. See Murray, “Dyes and fluorescent Whitening Agents for Paper,” Paper Chemistry 161-192 (2d ed. 1996). This lack of market penetration may be explained by the fact that pigments do not contain solubilizing functional groups and have little affinity for or substantivity to cellulose. In particular, the addition of a fixing agent, such as cationic starch, aluminum sulfate (alum) and cationic polymers, is typically required to fix pigments to cellulose fibers. Aluminum sulfate is the most common fixing agent for pigments and can also serve as an acidic sizing agent. However, neutral sizing agents have gained in popularity over acidic sizing agents, and aluminum sulfate can interfere with neutral sizing agents. An object of the invention is to provide a method for coloring cellulosic materials using an aqueous pigment dispersion which does not require fixing agents or alum. A feature of the method of the present invention is the use of a cationic dispersion containing at least one pigment, water, and at least one dispersing agent comprising a quaternary salt of a styrene maleimide imide resin to color cellulosic materials such as paper. An advantage of the method of the present invention is that it permits consistent coloring of cellulosic material over time, which is important in continuous and semi-continuous papermaking operations. Yet another advantage of the method of the present invention is that it exhibits essentially 100 percent, rapid exhaustion of the pigment particles into the cellulosic material, and thus generates clear backwaters. This is vitally important both from an economical and environmental vantage point. |
<SOH> SUMMARY OF THE INVENTION <EOH>In one aspect, the present invention relates to a method of coloring a cellulosic material, which includes a) dispersing pulped cellulosic material into water; and b) coloring the pulped cellulosic material by adding a cationic dispersion to the water, where the dispersion includes: (i) at least one pigment; (ii) water; and (iii) at least one quaternary salt of a styrene maleimide imide resin in an amount effective to disperse the pigment. In another aspect, the present invention relates to a colored cellulosic material, consisting essentially of pigment particles coated with a styrene maleimide imide resin; the coated particles fixed on fibers of a cellulosic material. In yet another aspect, the present invention relates to a cationic dispersion, which includes (i) at least one pigment; (ii) at least one dispersing agent comprising a quaternary salt of a styrene maleimide imide resin; and (iii) water. detailed-description description="Detailed Description" end="lead"? |
Method for coloring building materials using a cationic pigment dispersion |
A cationic dispersion which includes at least one pigment, water, and at least one quaternary salt of a styrene maleimide imide resin in an amount effective to disperse the organic pigment. A method for preparing the cationic dispersion includes (i) mixing, at 500 to 10,000 rpm, at least one pigment; water; and either (a) at least one a quaternary salt of a styrene maleimide imide resin or (b) at least one styrene maleimide imide resin in combination with at least one weak acid, thereby forming a dispersion premix; (ii) milling the dispersion premix in a mixer filled with ceramic, metal or glass beads for a period of time sufficient to reduce pigment agglomerates to primary particles, thereby forming a non-standardized dispersion; and (iii) standardizing the dispersion against a color standard by adding water. The resulting cationic dispersion exhibits good alkali resistance and lightfastness, and can be used to integrally color concrete and other building materials. |
1. A cationic dispersion, comprising: (i) at least one pigment; (ii) at least one dispersing agent comprising a quaternary salt of a styrene maleimide imide resin; and (iii) water. 2. The cationic dispersion of claim 1, wherein said pigment is an organic pigment is selected from the group consisting of phthalocyanine green, phthalocyanine blue, carbazole violet, toluidine red, Dalamar yellow, Watchung red and diketopyrrolopyrrole. 3. The cationic dispersion of claim 2, wherein said organic pigment is a phthalocyanine. 4. The cationic dispersion of claim 1, wherein said pigment is an inorganic pigment selected from the group consisting of red oxide, yellow oxide, black iron oxide, cobalt blue, carbon black and bismuth vanadate. 5. The cationic dispersion of claim 1, further comprising at least one member of the group consisting of a defoamer, a biocide and a viscosity control agent. 6. The cationic dispersion of claim 1, wherein said pigment comprises primary particles. 7. A process for preparing a cationic dispersion, comprising: (i) mixing, at 500 to 10,000 rpm, at least one pigment, water, and either (a) at least one a quaternary salt of a styrene maleimide imide resin or (b) at least one styrene maleimide imide resin in combination with at least one weak acid, thereby forming a dispersion premix; (ii) milling the dispersion premix in a mixer filled with ceramic, metal or glass beads for a period of time sufficient to reduce pigment agglomerates to primary particles, thereby forming a non-standardized dispersion; (iii) adding water to the non-standardized dispersion until it matches a color standard and forms a cationic dispersion suitable for coloring building materials. 8. The method of claim 7, wherein said pigment and water are mixed with at least one quaternary salt of a styrene maleimide imide resin. 9. The method of claim 7, wherein said pigment and said water are mixed with at least one styrene maleimide imide resin in combination with at least one weak acid. 10. The method of claim 9, wherein said weak acid is at least one member selected from the group consisting of acetic acid, citric acid, carbonic acid, hydrofluoric acid, oxalic acid and nitrous acid. 11. A colored building material, comprising the cationic dispersion of claim 1 dispersed in a building material. 12. The colored building material of claim 11, wherein said building material is selected from at least one member of the group consisting concrete, asphalt, plaster, mortar and cement mortar. |
<SOH> BACKGROUND OF INVENTION <EOH>1. Field of the Invention This invention relates to a method for coloring building materials such as concrete, asphalt, plaster, mortar and cement mortar. More particularly, this invention relates to a method for coloring building materials using a cationic dispersion which contains at least one pigment, water, and at least one dispersing agent comprising a quaternary ammonium salt of a styrene maleimide imide copolymer. 2. Description of the Prior Art It is desirable to color exposed concrete surfaces for both aesthetic and functional reasons. Colored concrete buildings do not present an environment as sterile and cold as white concrete. In addition, light colors can be used in sunny climates to help reduce glare while darker colors may be used to increase a building's heat storage capacity in cooler climates. Coloring the exposed surface by painting or by coating the surface with some other decorative layer is known. U.S. Pat. Nos. 2,549,516; 3,929,692 and 4,134,956 disclose compositions for covering exposed concrete surfaces. However, painting or application of a coating layer is an additional step in construction which adds cost and complexity to a construction project. Moreover, an exterior painted surface may require repainting in a relatively short period of time. Another approach is to add a color additive to the building material, thereby eliminating the extra step associated with painting the building surface. However, any color additive must be uniformly dispersed throughout the building material. This can be difficult given the low intensity mixing and short mixing times customary in the building materials industry. Moreover, the additive must not adversely affect the desirable properties of the building material, such as the strength or setting behavior of concrete or reduce the compressive strength or abrasion resistance of asphalt. See ASTM C 979-82 “Standard Specification for Pigments for Integrally Colored Concrete,” which contains some of the industry association standards for coloring concrete. Inorganic pigments are typically used as color additives for building materials and typically include iron oxides (natural and synthetic), chromium oxide, cobalt blue, and titanium dioxide. However, these inorganic pigments offer a limited range of colors and brightness. Organic pigments have not been used to color building materials as it is believed they lack sufficient alkali resistance and lightfastness. In 1981 the American Society for Testing and Materials (ASTM) in a report entitled “Pigments for Integrally Colored Concrete,” discussed the test results of various inorganic and organic pigments for lightfastness, alkali resistance, water wettability and curing stability. All of the organic pigments tested, including phthalocyanine green, failed to meet the lightfastness testing standards. Dry pigment powders have been used to color concrete compositions because they are highly dispersible. However, these powders have poor processing properties, and typically cake together and form lumps upon storage. They also tend to form dust. The use of free flowing granules or beads to overcome the processing problems and dust associated with dry pigment powders has been suggested. These granules may be produced by spray drying aqueous dispersions, as proposed by U.S. Pat. Nos. 4,946,505; 5,484,481; 5,853,476; and 5,882,395. However, the evaporation of the aqueous dispersion requires expensive equipment and significant energy expenditures which can make the use of such granules economically unattractive. Another approach is to modify the particle's surface to improve its dispersibility in aqueous solution. U.S. Pat. No. 5,401,313 discloses a pigment particle whose surface is coated with an electric charge modifying agent and a dispersion promoting agent. The dispersion promoting agent is selected from stearates, acetates, alkylphenols, cellulosics, waxes, lignins, acrylics, epoxies, urethanes, ethylenes, styrenes, propylenes and polymers having functions groups of alcohols, glycols, aldehydes, amides and carboxylic acids, and is preferably sodium lignosulfonate for cementitious application systems. The surface-modified particle may be produced in powder, dispersion or granular form, with bead granules having a particle size of 25 to 250 microns being preferred. Styrene maleimide imide resins and their quaternary salts are known. “Technical Information-Styrene Maleimide Resins SMA X 1000 I, X 2000 I, X 3000 I, X 4000 I,” Elf Atochem Brochure (1998) suggests their use in paper sizing, as a cationic dispersing resin for pigments and particles in acidic and cationic formulations, as cationic polymeric surfactants, and as cationic modifiers for waterbased coatings, inks and varnishes. However, there is no disclosure or suggestion concerning the use of these resins to disperse pigments in building materials. An object of the invention is to provide a method for coloring concrete using an aqueous pigment dispersion. A feature of the method of the present invention is the use of a cationic dispersion containing at least one pigment, water, and at least one dispersing agent comprising a quaternary salt of a styrene maleimide imide resin to color building materials such as concrete. An advantage of the present invention is the ability to color building materials such as concrete with bright organic pigments that do not suffer from poor alkali resistance and/or poor lightfastness. Another advantage of the present invention is that it permits the ready removal of graffiti or other surface defacement from a concrete surface without impairing its surface appearance. |
<SOH> SUMMARY OF THE INVENTION <EOH>In one aspect, the present invention relates to a cationic dispersion suitable for coloring building materials, which includes (i) at least one pigment; (ii) at least one dispersing agent comprising a quaternary salt of a styrene maleimide imide resin; and (iii) water. In another aspect, the present invention relates to a method for preparing a cationic dispersion suitable for coloring building materials, which includes (i) mixing, at 500 to 10,000 rpm, at least one pigment, water, and either (a) at least one a quaternary salt of a styrene maleimide imide resin or (b) at least one styrene maleimide imide resin in combination with at least one weak acid, thereby forming a dispersion premix; (ii) milling the dispersion premix in a mixer filled with ceramic, metal or glass beads for a period of time sufficient to reduce pigment agglomerates to primary pigment particles, thereby forming a non-standardized dispersion; (iii) adding water to the non-standardized dispersion until it matches a color standard and forms a cationic dispersion suitable for coloring building materials. detailed-description description="Detailed Description" end="lead"? |
Afm cantilevers and methods for making and using same |
The invention provides high performance cantilevers with optimal combinations of high resonant frequency and low force constant. In one aspect, AFM cantilevers with spring constants in the range 1-10−6 N/m with (fundamental) resonant frequencies in aqueous solutions of 0.1-100 MHz are provided. A high performance cantilever may be made by focused ion beam milling or electron deposition. The high performance cantilevers allow faster scanning, increase the temporal resolution of force measurement, improve measurement sensitivity by reducing cantilever noise, and improve sensitivity by reducing cantilever spring constant. |
1. A cantilever for use in a scanning probe microscope comprising a width to thickness ratio of about 3:1 or less and which is smaller in at least one dimension than about 5 μm. 2. A cantilever for use in a scanning probe microscope whose resonance frequency is reduced by less than 70% in solution compared to its resonance frequency in air. 3. A cantilever for use in a scanning probe microscope, wherein the body of the cantilever comprises a carbon nanotube. 4. The cantilever according to any of claims 1-3, wherein the cantilever has a resonant frequency equal to or above 10 kHz. 5. The according to any of claims 14, wherein the cantilever has a resonant frequency equal to or above 100 kHz. 6. The cantilever according to claim 1 or 2 wherein the cantilever comprises silicon, silicon nitride, silicon dioxide, a metal, a plastic, and a silicon-based rubber. 7. The cantilever according to claim 6, wherein the metal is selected from gold, aluminum, silver and nickel. 8. The cantilever according to claim 6, wherein the silicon-based rubber is PDMS. 9. The cantilever according to claim 1 or 2, wherein the cantilever comprises a reflective portion and/or comprises a conductive material. 10. The cantilever according to claim 1 or 2, wherein the cantilever comprises a width to thickness ratio of about 1:1 or less. 11. The cantilever according to any of claims 2-4, wherein the cantilever has at least one dimension which is smaller than 5 μm. 12. A method for producing a cantilever comprising: (a) providing a starting material; (b) exposing the starting material to an ion beam; and (c) removing molecules from the starting material to generate a cantilever which has a width to thickness ratio of about 3:1 or less and which is smaller in at least one dimension than about 5 μm. 13. A method for producing a high performance cantilever comprising: (a) providing a starting material; (b) exposing the starting material to an electron beam; and (c) depositing molecules on the starting material to generate a cantilever which has a width to thickness ratio of about 3:1 or less and which is smaller in at least one dimension than about 5 μm. 14. The method according to claim 12 or 13, further comprising imaging the starting material at one or more time intervals. 15. The method according to claim 12 or 13, wherein the starting material comprises silicon, silicon nitride, silicon dioxide, or a metal. 16. The method according to claim 12 or 13, wherein the cantilever comprises a resonant frequency above at least about 10 kHz. 17. The method according to claim 12 or 13, wherein the cantilever comprises a resonant frequency above at least about 100 kHz. 18. The method according to claim 12 or 13, wherein the cantilever comprises a spring constant of at least about 1-10−6 N/m. 19. The method according to claim 12 or 13, wherein the starting material is a beam, a film, a sheet, a V-shaped material, or a rectangular shaped material. 20. The method according to claim 12 or 13, wherein the starting material is a cantilever. 21. The method according to claim 12 or 13, further comprising a step of generating a tip at an end of the cantilever. 22. The method according to claim 12 or 13, wherein the tip is generated by electron beam deposition. 23. The method according to claim 12 or 13, wherein the cantilever comprises a reflective surface. 24. The method according to claim 12 or 13, wherein the starting material is a conductive material. 25. A method of measuring a property of a cantilever, comprising measuring deflection of a cantilever according to claims 1-3. 26. A method for measuring a property of a sample, comprising: detecting an interaction between a cantilever according to any of claims 1-3 and the sample, wherein the interaction provides a measure of the property of the sample. 27. The method according to claim 26, wherein the property comprises one or more surface features of the molecule. 28. The method according to claim 26, wherein the cantilever further comprises one or more biological molecules and wherein the one or more biological molecules interact with one or more molecules of the sample. 29. The method according to claim 28, wherein the biological molecule binds to the one or more molecules. 30. The method according to claim 28, wherein the one or more biological molecules is selected from the group consisting of: nucleic acids, proteins, polypeptides, peptides, receptors, ligands, enzymes, antigens, drug molecules, therapeutic agents, lipids, lipid bilayers, detergents, a cell membrane fraction, organelles, and zwitterions. 31. The method according to claim 28, wherein the nucleic acid is selected from the group consisting of: a DNA molecule, RNA molecule, antisense molecule, ribozyme, triple helix forming molecule, an aptamer; and combinations and modified forms thereof. 32. The method according to claim 31, wherein the sample comprises one or more a cell, nucleic acids, proteins, polypeptides, peptides, receptors, ligands, enzymes, antigens, drug molecules, therapeutic agents, lipids, a cell membrane fraction, organelles, and microorganisms. 33. The method according to claim 31, wherein the property of the sample comprises one or more of: surface topography, binding, a chemical reaction, a cellular response, or polymerization. 34. A method for fabricating a nanostructure comprising: providing a substrate comprising a plurality of molecules; using a cantilever according to any of claims 1-3 to move one or more molecules on the substrate to a desired position on the substrate. 35. The method according to claim 34, wherein the cantilever is used to create a data structure on the substrate. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Scanning probe microscopes (SPMs) obtain data regarding surface topography by using a sharp tip or probe on the end of a cantilever held on or at a short distance (e.g., about 5-500 Å) from a sample. The cantilever tip can be deflected by various forces acting at the interface between the sample and the tip, such as electrostatic, magnetic, and van der Waals forces. A movement of the cantilever due to interactions between an atom at the end of the tip and an atom of the sample can then be measured electrically (as in a Scanning Tunneling Microscope, or STM) or optically (as in an Atomic Force Microscope, or AFM). By scanning the sample in x- and y-directions to change its position relative to the cantilever tip, three-dimensional information regarding the surface features of a sample can be obtained (see, e.g., Binnig, et al, 1986 , Phys. Rev. Lett. 56(9): 930-933); McClelland, et al., 1987, Rev. Progr. Quart. Non - Destr. Eval. 6: 1307; Martin, et al, 1987 , J. Appl. Phys. 61(10): 4723-4729). AFM cantilevers operate in an oscillating mode or in a non-oscillating mode and can further interact with a sample in a contact mode or in a non-contact mode. In an oscillating contact mode, the cantilever is oscillated mechanically at or near its resonant frequency so that its tip repeatedly taps a sample surface, thus reducing the tip's oscillation amplitude. In an oscillating non-contact mode, interactions between the sample and the tip alter the tip's oscillation amplitude or frequency. The change in oscillation amplitude indicates proximity to the sample surface and may be used as a signal for feedback (e.g., for control of probe scanning). In a non-oscillating contact mode, the cantilever is not oscillated, and cantilever deflection is monitored as the tip is dragged over the sample surface, while in a non-oscillating non-contact mode, attractive interactions between the tip and the sample shift the cantilever resonance frequency. Atomic force microscopy is emerging as an important tool in methods which rely on detecting information about surface features of a sample, measuring forces between two surfaces, or fabricating nanostructures (e.g., on silicon wafers, thin film magnetic read/write heads, and the like) (see, e.g., U.S. Pat. No. 6,337,479). Atomic force microscopy also has many applications in biomedical research. It can be used for high contrast, high resolution imaging of biological surfaces in a wide range of environments (Engel, et al., 1999, Trends Cell Biol. 9: 77-80; Czajkowsky and Shao, 1998 , FEBS Lett. 430: 51-4.1998; Bustamante, et al., 1997, Curr. Opin. Struc. Biol. 7: 709-16; Hansma and Hoh, 1994 , Ann. Rev. Biophys. Biomol. Struct. 23: 115-39.) It also can be used to measure intermolecular forces (e.g., Heinz and Hoh; 1999 , Trends Biotech. 17:143-150; Mann, S. and H. E. Gaub, 1997 , Curr. Opin. Colloid Interface Sci. 2: 145-152; Cappella and Dietler, 1999 , Surf. Sci. Rep. 34: 1), intramolecular forces (Lee, et al., 1994; Lee, et al., 1994, Science 266: 771-773. 1994; Rief, et al., 1997 , Science 275: 1295-7), and local mechanical properties (A-Hassan et al., 1998, Biophys. J. 74: 1564-1578; Vinckier and Semenza, 1998 , FEBS Lett. 430: 12-6. 1998; Radmacher, et al., 1996, Biophys. J. 70: 556-567). Imaging with AFM offers advantages for studying biological samples, because the samples do not require drying, sectioning, metal coating or chemical fixing prior to analysis. Thus, AFMs may be used with samples that require very little sample preparation, including samples that are biologically active in both ambient air (including dried samples) and liquid. One of the limiting elements in current AFMs is the design of the cantilever. The performance of the cantilever is primarily constrained by a combination of its fundamental resonant frequency (ω) and spring constant (k). Typical cantilevers of the prior art are on the order of 85-500 μm long and have resonant frequencies substantially less than 500 KHz. For example, generally, prior art cantilevers have lengths on the order of 85-320 μm, widths of 10-20 μm and thicknesses on the order of 0.5 μm. This produces typical [k, ω s ] pairs of [0.5 N/m, 30 kHz] for shorter cantilevers, and [0.01 N/m, 2 kHz] for the longer cantilevers (where ω s is the fundamental resonant frequency in solution). Smaller cantilevers with higher resonant frequencies are desirable because they allow faster imaging rates and permit the cantilever tip to more closely track sample topography (see, e.g., Butt, Biophys. J. 60: 777-785). Smaller cantilevers also are less affected by viscous damping, and are therefore more sensitive (see, e.g., U.S. Pat. No. 6,016,693). Smaller cantilevers have been described in Walters, et al., 1996, Rev. Sci. Instrum. 67: 3583-3590 (23 μm length); Walters, et al., 1997, SPIE, Proceedings Micro - Machining and Imaging 3009: 48 (26 μm length); and Schaeffer, et al., 1997, SPIE, Proceedings Micro - Machining and Imaging 3009: 48 (9 μm length). A number of groups have made efforts to generate high performance cantilevers. Stowe, et al., 1997, Appl. Phys. Lett. 7(1): 288-290, describe ultrathin (60 nm) silicon cantilevers with force constants on the order of 10 −6 and resonant frequencies in vacuum of 1.7 kHz. While the performance of these cantilevers in solution was not examined, the dimensions of the cantilevers are such that they have a predicted resonant frequency in water of about 200 Hz. This frequency is too low to be useful in most biological applications. Ried, et al., 1997, J. Microelectromechanical Sys. 6: 294-302, describe piezo resistive cantilevers with resonant frequencies of 6 MHz and spring constants of 2 N/m. However, these types of cantilevers have relatively poor detection sensitivity and are difficult to work with in solution, preventing them from being used productively in biological research. Such cantilevers are used instead in data storage applications (see, e.g., Mamin and Rugar, 1996 , Appl. Phys. 79: 5644-5644) or in force-based magnetic resonant imaging (Rugar et al., 1994, Science 264: 1560-1563). High performance cantilevers specifically for use in biological research have been described (Walters et al., 1996, Rev Sci Instrum 67:3583-3590; Viani, et al., 1999, Rev. Sci. Instrum. 70: 4300-4303). The best cantilevers thus far developed have resonant frequencies of 100-200 kHz in solution and spring constants of 0.1-0.2 N/m. However, improvements in these cantilevers are largely limited by the lithographic and thin film deposition methods used for their fabrication. U.S. Pat. No. 6,016,693 discloses a method for making a smaller cantilever (e.g., 2-10 μm in length). The method comprises fabricating a “sacrificial cantilever” of SiO 2 and depositing a layer of material which will form the final cantilever onto the sacrificial cantilever. The sacrificial cantilever is then etched away. U.S. Pat. No. 5,666,190 discloses a compound cantilever for a scanning probe microscope which includes a bending portion and a vibrating portion. The vibrating portion has a lower mechanical resonant frequency than the bending portion. The cantilever is fabricated from two fused silicon oxide wafers. |
<SOH> SUMMARY OF THE INVENTION <EOH>The invention provides high performance cantilevers for Atomic Force Microscopes (AFMs), as well as methods for making and using the same. In one aspect, the invention provides a cantilever with resonant frequencies in the range of 1-100 MHz in solution for cantilevers with a spring constant of 0.1 N/m. At the high end, this resonant frequency is approximately one to three orders of magnitude better than the best cantilevers that are currently available. In one aspect, the invention provides a cantilever for use in a scanning probe microscope comprising a width to thickness ratio of about 3:1 or less and which is smaller in at least one dimension than about 5 μm. More preferably, the cantilever comprises a width to thickness ratio of about 1:1 or less. In another aspect, the invention provides a cantilever for use in a scanning probe microscope whose resonance frequency is reduced by less than 70% in solution compared to its resonance frequency in air. In a further aspect, the invention provides a cantilever for use in a scanning probe microscope, wherein the body of the cantilever comprises a carbon nanotube. Preferably, the cantilevers have a resonant frequency equal to or above 10 kHz. More preferably, the cantilevers have a resonant frequency equal to or above 100 kHz. Cantilevers may be fabricated from a variety of materials, including, but not limited to silicon, silicon nitride, silicon dioxide, a metal (e.g., gold, aluminum, silver, or nickel), a plastic, and a silicon-based rubber (e.g., PDMS). The metal gold, aluminum, silver and nickel. In one aspect, the cantilever comprises a reflective portion. In another aspect, the cantilever comprises a conductive material. The invention also provides a method for producing a cantilever comprising providing a starting material; exposing the starting material to an ion beam; and removing molecules from the starting material to generate a cantilever which has a width to thickness ratio of about 3:1 or less and which is smaller in at least one dimension than about 5 μm. The invention further provides a method for producing a high performance cantilever comprising: providing a starting material; exposing the starting material to an electron beam; and depositing molecules on the starting material to generate a cantilever which has a width to thickness ratio of about 3:1 or less and which is smaller in at least one dimension than about 5 μm. In one aspect, the methods further comprise the step of imaging the starting material at one or more time intervals. The starting material may comprise silicon, silicon nitride, silicon dioxide, or a metal. The method can be used to generate cantilevers comprising a resonant frequency above at least about 10 kHz, and preferably, above at least about 100 kHz. The cantilever may comprise a spring constant of about 1-10 −6 N/m. Starting materials may comprise materials of a variety of shapes. For example, the starting material may be a beam, a film, a sheet, a V-shaped material, or a rectangular shaped material. In one aspect, the starting material is itself a cantilever. In one aspect, a tip is generated at the end of the cantilever. For example, a tip may be fabricated using electron beam deposition. In one aspect, the cantilevers produced by the methods described above comprise a conductive material. The invention also provides a method of measuring the property of a cantilever comprising measuring the deflection of any of the cantilevers described above. The invention further provides a method for measuring a property of a sample, comprising: detecting an interaction between a cantilever according to the invention and a sample, wherein the interaction provides a measure of the property of the sample. The property may comprise one or more surface features of the sample. In one aspect, the cantilever further comprises one or more biological molecules and the one or more biological molecules interact with one or more molecules of the sample. In another aspect, the biological molecule binds to the one or more molecules. In a further aspect, the one or more biological molecules is selected from the group consisting of: nucleic acids, proteins, polypeptides, peptides, receptors, ligands, enzymes, antigens, drug molecules, therapeutic agents, lipids, lipid bilayers, detergents, a cell membrane fraction, organelles, and zwitterions. The sample may comprise one or more of: a cell, nucleic acids, proteins, polypeptides, peptides, receptors, ligands, enzymes, antigens, drug molecules, therapeutic agents, lipids, a cell membrane fraction, organelles, and microorganisms. The invention additionally provides a method for fabricating a nanostructure comprising: providing a substrate comprising a plurality of molecules; and using a cantilever as described above to move one or more molecules on the substrate to a desired position on the substrate. In one aspect, the cantilever is used to create a data structure on the substrate. |
Desalting plate for maldi mass spectrometry |
A novel Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) desalting sample support plate is described. The plate comprises a sample presentation surface, wherein the sample presentation surface comprises at least one receiving surface of an absorbent layer, wherein the absorbent layer retains selected molecules on the receiving surface. Methods for making and using the sample support plate in conventional and automated MALDI-MS are also described. |
1. A desalting sample support plate, for use in Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS), comprising a sample presentation surface, wherein the sample presentation surface comprises at least one receiving surface of an adsorbent layer, wherein the adsorbent layer retains selected molecules on the receiving surface. 2. The sample plate of claim 1, wherein the adsorbent layer is sufficiently adsorbent to allow retention of selected molecules on the adsorbent layer. 3. The sample plate of claim 1, wherein the adsorbent layer is washed with an aqueous solution to remove the salts from the adsorbent layer. 4. The sample plate of claim 3, the selected molecules that are retained on the surface of the adsorbent layer are selected from the group consisting of an analyte, a matrix, and a combination thereof. 5. The sample plate of claim 2, wherein the adsorbent layer is a polymeric film. 6. The sample plate of claim 5, wherein the polymeric film is prepared by admixing a monomer, a polymer, or a combination thereof, a porogen, and an initiator. 7. The sample plate of claim 5, wherein the polymeric film is prepared by admixing a monomer, a polymer, or a combination thereof, and an initiator. 8. The method of claim 6 wherein the polymer is selected from the group consisting of polystyrene and polyvinylpyrrolidinone. 9. The sample plate of claim 6, wherein the monomer is selected from the group consisting of a monovinyl monomer, a polyvinyl monomer, and a mixture of monovinyl and polyvinyl monomers. 10. The sample plate of claim 8, wherein the monovinyl monomer is selected from the group consisting of styrene, N-vinylpyrrolidone, methacrylate, vinylacetate, glycidyl methacrylate, and any combination thereof. 11. The method of claim 8, wherein the monovinyl monomer is N-vinylpyrrolidone. 12. The sample plate of claim 8, wherein the polyvinyl monomer is selected from the group consisting of divinylbenzene, ethylene dimethacrylate, bis-acrylamide, divinylpyridine, ethylene dimethacrylate, hydroxyalkylene dimethacrylate, and any combination thereof. 13. The method of claim 8, wherein the polyvinyl monomer is divinylbenzene. 14. The method of claim 8, wherein the mixture of monovinyl monomer and polyvinyl monomer is divinylbenzene and N-vinylpyrrolidone. 15. The sample plate of claim 6, wherein the porogen is selected from the group consisting of aliphatic hydrocarbons, aromatic hydrocarbons, esters, alcohols, ketones, ether, and any combination thereof. 16. The sample plate of claim 6, wherein the initiator is selected from the group consisting of benzoyl peroxide, lauroyl peroxide, peroxodisulfate, Vazo 52, Vazo 64, Vazo 67, Vazo 88, V70, and any combination thereof. 17. The MALDI-MS sample plate of claim 1, wherein the receiving surface is in a precisely defined location. 18. The MALDI-MS sample plate of claim 12, wherein the precisely defined location of the receiving surface facilitates automated analysis. 19. The MALDI-MS sample plate of claim 17, comprising a plurality of receiving surfaces, such that a grid of receiving surfaces is formed on the sample presentation surface. 20. The MALDI-MS sample plate of claim 19, wherein the grid comprises 96 receiving surfaces. 21. The MALDI-MS sample plate of claim 19, wherein the plurality of receiving surfaces facilitates high throughput analysis. 22. A method for preparing a sample for Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) comprising: providing the sample support plate of claim 1; applying a sample to the receiving surface; allowing selected molecules in the sample to be retained on the adsorbent layer; and applying a solvent to the receiving surface to wash away salts and other impurities, thereby desalting the sample applied to the receiving surface; to thereby prepare a sample for MALDI-MS. 23. The method of claim 22, wherein prior to applying the sample to the receiving surface, the sample support plate is treated with a polar organic solvent. 24. The method of claim 23, wherein the polar organic solvent is selected from the group consisting of acetonitrile, methanol, and water/organic solvent mixtures. 25. The method of claim 24, wherein the polar organic solvent is acetonitrile. 26. The method of claim 22, wherein the adsorbent layer is washed with an aqueous solution to remove the salts from the adsorbent layer. 27. The method of claim 22, wherein the sample comprises an analyte of interest, a matrix material, one or more salts, and one or more solvents. 28. The method of claim 27, wherein the solvent is aqueous. 29. The method of claim 22, wherein the sample contains a solution of a matrix material. 30. The method of claim 22, wherein the sample does not contain a matrix material. 31. The method of claim 30, wherein the matrix material is added after the adsorbent layer is washed with an aqueous solution. 32. A method for preparing the sample support plate of claim 1 comprising: providing a sample support plate comprising a sample presentation surface; forming on the top sample presentation surface at least one receiving surface of an adsorbent layer, wherein the adsorbent layer retains selected molecules on the receiving surface. 33. A method for preparing the sample support plate of claim 1 comprising: forming, on a sample support plate having a sample presentation surface, at least one receiving surface of an adsorbent layer, wherein the adsorbent layer retains selected molecules on the receiving surface. 34. The method of claims 32, wherein the adsorbent layer comprises a polymeric film. 35. The method of claim 34, further comprising admixing a monomer, a polymer, or a combination thereof, a porogen, and an initiator; coating the target spot with the admixture; initiating a polymerization reaction to form a polymeric film; and washing the polymeric film to remove residual monomer, porogen and initiator. 36. The method of claim 34, further comprising admixing a monomer, a polymer, or a combination thereof, and an initiator; coating the target spot with the admixture; initiating a polymerization reaction to form a polymeric film; and washing the polymeric film to remove residual monomer or polymer, porogen, and initiator. 37. The method of claim 35, further comprising treating the polymer film with a polar organic solvent. 38. The method of claim 37, wherein the polar organic solvent is selected from the group consisting of acetonitrile, methanol, and water/organic solvent mixtures. 39. The method of claim 38, wherein the polar organic solvent is acetonitrile. 40. The method of claim 35, wherein the polymer is selected from the group consisting of polystyrene and polyvinylpyrrolidinone. 41. The method claim 35, wherein the monomer is selected from the group consisting of a monovinyl monomer, a polyvinyl monomer, and a mixture of monovinyl and polyvinyl monomers. 42. The method of claim 41, wherein the monovinyl monomer is selected from the group consisting of styrene, N-vinylpyrrolidone, methacrylate, vinylacetate, glycidyl methacrylate, and any combination thereof. 43. The method of claim 41, wherein the monovinyl monomer is N-vinylpyrrolidone. 44. The method of claim 41, wherein the polyvinyl monomer is selected from the group consisting of divinylbenzene, ethylene dimethacrylate, bis-acrylamide, divinylpyridine, ethylene dimethacrylate, hydroxyalkylene dimethacrylate, and any combination thereof. 45. The method of claim 41, wherein the polyvinyl monomer is divinylbenzene. 46. The method of claim 41, wherein the mixture of monovinyl monomer and polyvinyl monomer is divinylbenzene and N-vinylpyrrolidone. 47. The method of claim 35, wherein the porogen is selected from the group consisting of aliphatic hydrocarbons, aromatic hydrocarbons, esters, alcohols, ketones, ether, and any combination thereof. 48. The method of claim 47, wherein the initiator is selected from the group consisting of benzoyl peroxide, lauroyl peroxide, peroxodisulfate, Vazo 52, Vazo 64, Vazo 67, Vazo 88, V70, and any combination thereof. 49. A method for performing Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) on an analyte of interest, comprising: providing a sample support plate comprising a sample presentation surface, wherein the sample presentation surface comprises at least one receiving surface of an adsorbent layer, wherein the adsorbent layer retains selected molecules on the receiving surface; applying a sample comprising an analyte of interest to the receiving surface of the adsorbent layer; allowing the analyte of interest in the sample to be retained on the adsorbent layer; washing the adsorbent layer with an aqueous solution to remove the salts from the adsorbent layer, thereby desalting the sample; and performing MALDI-MS on the desalted analyte of interest retained on the adsorbent layer. 50. A plate or method of claim 1, wherein the presentation surface comprises at least one sample target spot, wherein the target spot comprises the adsorbent layer. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Mass spectrometry (MS) with ionization by matrix-assisted laser desorption and ionization (MALDI) has become a useful tool for the analysis of large molecules, e.g., biopolymers, such as proteins, peptides, oligonucleotides, DNA, RNA, etc. It is well known that the sensitivity of the analysis and the speed of automation are highly dependent on preparation and purity of the sample on the MALDI plate. Many of the issues associated with sample preparation for MALDI mass spectrometry are summarized in Bruker's UK patent application (GB 2332273A). In particular, salts can dramatically affect the quality of the resulting mass spectra due to adduct formation. One existing method for salt removal utilizes reversed-phase sorbents such as C18-silica or divinylbenzene-based polymers. For example, a mixture of analytes is passed through a packed bed containing such sorbents, and the analytes adsorb to the packing material, while the salts are not retained. The adsorbed analytes are subsequently eluted from the sorbent with an appropriate solvent. Several commercially available products are available for this application. However, all of these products require several manipulation steps prior to application of the sample onto the MALDI plate. These additional steps may lead to significant sample loss and potential re-contamination of the sample. In addition, samples typically become diluted during the sample preparation step because of the relatively large elution volumes that are required. Because the MALDI laser focuses on only a small percentage of the total area of the applied sample, only a small fraction of the applied diluted sample will be vaporized and detected, resulting in decreased sensitivity. An additional problem with spotting a sample onto a traditional MALDI plate pertains to the drying of the sample on the plate. As the sample dries, the target analyte randomly concentrates in localized regions. As a result, analysis time for each sample increases, as the laser may need to search until it finds the target analyte that is sufficient for analysis. For example, when a drop of sample and matrix solution that is placed onto a clean metal sample support plate dries on a metal surface, the sample spot consisting of small matrix crystals spreads over the formerly wet area. In general, the wetted area is not uniformly coated. In aqueous solutions, most of the small crystals of the matrix generally begin to grow at the margin of the wet area on the metal plate and continue to grow toward the center of the wet area. Thus the analyte molecules are irregularly distributed, and the center of the spot is frequently empty or covered with fine small crystals. These crystals are not typically sufficient for MALDI ionization due to the high concentration of salts. Furthermore, the MALDI ionization yield and mass resolution fluctuate in the sample spot from site to site. It is often a troublesome process to find a favorable location on the sample spot with good analyte ion yield and good mass resolution. Consequently, the development of high sample throughput automation of MALDI mass spectrometry analysis has been hindered. British patent application GB 2332273A describes a MALDI plate, coated with a Teflon-like hydrophobic coating having hydrophilic patches (“anchors”), which utilizes surface property (hydrophilic or hydrophobic) modification on the plate. After sample droplets are deposited onto the anchors, the droplets shrink during solvent evaporation, thereby centering themselves onto the anchor positions. Thus, MS detection sensitivity increases 10 to 100 times as compared to the conventional dried sample droplet preparation method described above, because the analyte is concentrated in smaller spots. The sample spots can be arranged in a precise grid to facilitate rapid, automated MALDI-MS. Such coated plates (AnchorChip™) are marketed by Bruker Daltonics®. However, these plates are incapable of retaining analyte molecules during a wash step, used to remove salts that may interfere with the analysis of the analyte. U.S. Pat. Nos. 6,020,208 and 6,124,137 describe a MALDI-MS plate design in which the presentation surface of the target spot is derivitized with affinity molecules which allow for specific or nonspecific capturing of analytes of interest on the presentation surface. The sensitivity as well as the efficiency of the analysis by MALDI-MS are reportedly improved over conventional methods. The molecules that are not used for analysis may be subjected to further processing and subsequently further analysis on the presentation surface. Published PCT applications WO 98/59360, WO 98/59361, WO 98/59362, and their corresponding European patent applications, EP 00990256, EP 00990257, and EP 00990258 describe plates, and methods of retentate chromatography for retaining selected molecules on a variety of adsorbents and using a variety of selectivity conditions. Upon selection of the highest affinity conditions direct MALDI-MS is used to analyze the selected molecule. U.S. Pat. No. 5,480,526 describes the removal of salts from samples analyzed by MALDI-MS by using capillary electrophoresis methods prior to the analysis. U.S. Pat. No. 6,104,028 describes the improvement of the sensitivity and resolution of MALDI-MS by improvement of the matrices. U.S. Pat. Nos. 5,260,571, 5,281,538, and 5,308,978 describe improvement in sensitivity and resolution of detection by applying the matrix as a layer to the target spot of the MALDI-MS sample plate prior to the application of the sample, to thereby provide, after drying, an intimate mixture of sample and matrix material. Similarly, U.S. Pat. No. 5,595,636 describes the use of a thin lacquer-like smooth matrix layer applied prior to the application of the sample to assist in the desorption of the analyte molecules. U.S. Pat. Nos. 5,770,860, 5,770,272, and 5,705,813 describe improved devices for use in connection with mass spectrometric analysis. In particular, U.S. Pat. No. 5,770,272 describes a nebulizing sprayer that deposits a continuous, homogeneous layer of MALDI matrix material on MALDI targets. U.S. Pat. No. 5,705,813 describes an integrated liquid handling system for manipulation of a sample prior to mass analysis by MALDI-MS. U.S. Pat. No. 5,770,860 discloses a method for loading sample supports of a mass spectrometer using a multiple pipette unit to simultaneously transfer multiple samples from microtiter plates. Repeated loading, in this fashion, generates high-density samples on the sample plate. Nevertheless, a need exists for technological advancement of the MALDI-MS sample design that is capable of desalting samples directly on the MALDI plate while retaining the analyte and matrix on an underivatized surface, such that the salts may be washed away by the use of an appropriate solvent, e.g., water, and thereby reduce the number of sample manipulation steps while improving the sensitivity and resolution of detection by MALDI-MS by MALDI-MS. The use of small spots of the sample would localize the sample into a specific area, removing the requirement of searching for the sample location. |
<SOH> SUMMARY OF THE INVENTION <EOH>The invention is directed to a desalting sample support plate for MALDI-MS. The invention provides convenient methods of preparation and use of the support plate. Additionally, the invention provides methods of sample preparation and analysis. Furthermore, the methods of sample preparation and analysis of the present invention are capable of desalting samples directly on the MALDI plate while retaining the analyte and matrix on an underivatized surface, such that the salts may be washed away by the use of an appropriate solvent, e.g., water, and thereby reduce the number of sample manipulation steps while improving the sensitivity and resolution of detection by MALDI-MS. The use of small spots of the sample localizes the sample into a specific area, thereby obviating the need to search for the sample location. Thus, in one aspect, the invention is a desalting sample support plate for use in Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS). The MALDI plate comprises a sample presentation surface, wherein the sample presentation surface comprises at least one receiving surface of an adsorbent layer. The adsorbent layer retains selected molecules on the receiving surface. Another aspect of the invention is a method for preparing a sample for MALDI-MS. Providing the sample support plate described above, a sample is applied to the receiving surface. Selected molecules in the sample are retained on the adsorbent layer. A solvent is applied to the receiving surface to wash away salts and other impurities, thus desalting the sample applied to the target spot, to thereby prepare a sample for MALDI-MS. In yet another aspect, the invention is a method for preparing the sample support plate described above. A sample support plate is provided, comprising a sample presentation surface, and at least one receiving surface of an adsorbent layer. The adsorbent layer retains selected molecules on the receiving surface. In a related aspect, the invention is a method for preparing the sample support plate as described above. The method comprises forming, on a sample support plate having a sample presentation surface, at least one receiving surface of an adsorbent layer. The adsorbent layer retains selected molecules on the receiving surface. Another aspect of the invention is a method for performing Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) on an analyte of interest using the desalting plate described above. The method comprises providing a sample support plate comprising a sample presentation surface, wherein the sample presentation surface comprises at least one receiving surface of an adsorbent layer, wherein the adsorbent layer retains selected molecules on the receiving surface; applying a sample comprising an analyte of interest to the receiving surface of the adsorbent layer; allowing the analyte of interest in the sample to be retained on the adsorbent layer; washing the adsorbent layer with an aqueous solution to remove the salts from the adsorbent layer, thereby desalting the sample; and performing MALDI-MS on the desalted analyte of interest retained on the adsorbent layer. |
Method for the biochemical detection of analytes |
The invention relates to a method for detecting and/or quantifying analytes from a sample on an analysis carrier that has been formatted using a digital data code. Detection fields comprising the sensor elements required for the respective detection process, together with additional data structures in a defined digital format, are provided on the analysis carrier and combined to form sequcnces of formatted structures that can be interpreted as code words. To detect and quantify an analyte in a sample, the latter is applied to the analysis carrier and the formation of signal-generating elements is initiated at locations of molecular interaction. The localisation of signal-generating elements in the respective detection fields causes a formatted structure at this location to be replaced by another. This leads to the conversion of one code word into another within the predetermined quantity of valid code words. Both code words can be sequentially read and interpreted in the predetermined format. The statement concerning a successful or unsuccessful reaction is based on a comparison of the respective code words prior to and after detection. Detection takes place using a reading device, which is preferably constructed from components of the consumer goods industry. |
1. A method for detecting and/or quantifying at least one analyte in a sample on an analysis support, at least one defined sequence of fields being applied to the surface of the analysis support, characterized in that a subset of the fields constitute detection fields; a signal of one type is present on each field; signals of at least one defined sequence of fields can be interpreted as a digital codeword; sensor elements are applied to the detection fields in a controlled way; analytes are brought in contact with the analysis support for the purpose of molecular interaction with the sensor elements on the detection fields; signaling elements are localized on the detection fields when a molecular interaction has taken place; one type of signal on the detection field where the molecular interaction has taken place is replaced by another type of signal in a predetermined way by the localization of signaling elements; after the replacement of a signal of one type by a signal of another type on at least one detection field within a defined sequence of fields, the interpretation of this sequence of fields gives a different codeword than before the molecular interaction; comparison of the codeword read after the detection with the known codeword before the detection gives the detection result. 2. The method as claimed in claim 1, characterized in that any replacement of a signal of one type by a signal of another type on a non-detection field within a defined sequence of fields is identified as an error during the interpretation. 3. The method as claimed in one of the preceding claims, characterized in that the signals on the fields can be read and interpreted in a format known from digital storage technology. 4. The method as claimed. in one of the preceding claims, characterized in that the signaling elements are designed and localized so that they can be read and interpreted in a format known from digital storage technology. 5. The method as claimed in one of the preceding claims, characterized in that a certain number of codewords have no detection fields and are used for separating and/or addressing sizable code blocks. 6. The method as claimed in one of the preceding claims, characterized in that the sensor elements of one type can be unequivocally assigned to at least one defined detection field on the analysis support, and vice versa. 7. The method as claimed in one of the preceding claims, characterized in that the fields on the analysis support a shaped as spots, strips, circles or spirals, or have another geometrical shape. 8. The method as claimed in one of the preceding claims, characterized in that individual fields on the analysis support are arranged in the form of a spot matrix or a circular, spiral, strip-shaped, linear or other geometrical or stochastic structure. 9. The method as claimed in one of the preceding claims, characterized in that sequences of defined fields which represent codewords are arranged successively in the form of a track on the analysis support. 10. The method as claimed in one of the preceding claims, characterized in that the tracks are arranged circularly, spirally, linearly or in another defined way on the analysis support. 11. The method as claimed in one of the preceding claims, characterized in that biologically active substances such as sugars, steroids, hormones, lipids, proteins, in particular monoclonal or polyclonal or recombinant antibodies, peptides, antigens of any type, haptens, DNA, RNA, as well as natural and artificial derivatives thereof, in particular aptamers and PNA, organic-chemical active agent libraries, cells, microorganisms, viruses or parts thereof, preparations and extracts from biological materials, metabolites and the like can be used as the sensor elements. 12. The method as claimed in one of the preceding claims, characterized in that any resonant processes such as absorption, fluorescence, phosphorescence, plasmon resonance, quenching etc., and nonresonant processes such as reflection, diffraction, scattering etc., from spectroscopy can be used to generate the signals. 13. The method as claimed in one of the preceding claims, characterized in that electromagnetic effects such as piezo, resonance shift, capacitance change, Hall effect, magnetic effects, electrical charge displacement etc. can be used to generate the signals. 14. The method as claimed in one of the preceding claims, characterized in that microspheres of any shape and size, such as metal, magneto, silica or fluorescence-labeled beads, fluorescent or radioactive labels as well as molecular complexes or aggregates, layers of precipitates, or dyestuffs can be used as signaling elements. 15. The method as claimed in one of the preceding claims, characterized in that biological objects such as cells, bacteria, pollens, virus particles or parts thereof can be used as signaling elements. 16. The method as claimed in one of the preceding claims, characterized in that bodies and coatings, in particular metal grains, are formed as signaling elements on an initiator, in particular an electron donor, coupled to analyte molecules, at the site of the interaction. 17. The method as claimed in one of the preceding claims, characterized in that a binding, polymerization, precipitation, deposition or color reaction, or other chemical or biological reactions, are used to form signaling elements. 18. The method as claimed in one of the preceding claims, characterized in that the signals generated by the signaling elements are digitized by means of a threshold criterion. 19. The method as claimed in one of the preceding claims, characterized in that the dimensions of the signaling elements can be adapted to the dimensions of the detection fields. 20. The method as claimed in one of the preceding claims, characterized in that the signaling elements are designed so that one and only one signaling element of is localized on each detection field. 21. The method as claimed in one of the preceding claims, characterized in that the fields and the signaling elements can have dimensions smaller than 10 μm, preferably smaller than 2 μm and in particular smaller than 1 μm. 22. The method as claimed in one of the preceding claims, characterized in that the analyte in the sample is quantified with the aid of calibration fields, defined threshold criteria and/or statistics via multiple determination. 23. The method as claimed in one of the preceding claims, characterized in that a synchronization track with standardized substances, surface coatings or other structures for calibrating the reader and the detection is applied to the analysis support. 24. The method as claimed in one of the preceding claims, characterized in that standard substances for positive or negative controls are applied to defined detection fields and/or to adjacently or successively arranged rows of such detection fields. 25. The method as claimed in one of the preceding claims, characterized in that different concentrations are sensor elements of one type are applied to defined detection fields and/or to adjacently or successively arranged rows of such detection fields. 26. The method as claimed in one of the preceding claims, characterized in that any desired combination of conventional data stores, such as magnetic strips, card chips, barcodes, CD-ROM or CD-R, are integrated on the analysis support. 27. The method as claimed in one of the preceding claims, characterized in that the software, databases, signatures of other information with any desired configuration is present on the analysis support. 28. The method as claimed in one of the preceding claims, characterized in that encoding or identification of the detection to be carried out on the analysis support is present on the analysis support, in the same format or in another separately applied format. 29. The method as claimed in one of the preceding claims, characterized in that the fields are designed and arranged so that they can be read by means of a commercially available barcode reader. 30. The method as claimed in one of the preceding claims, characterized in that the analysis support is comparable in its physical features, such a shape, material, optical density and material thickness, as well as handling, to a magnetic card known from mass storage technologies. 31. The method as claimed in one of the preceding claims, characterized in that the analysis support is comparable in its physical features, such a shape, material, optical density and material thickness, as well as handling, to a CD, CD-ROM or DVD known from mass storage technologies, or successors thereof. 32. The method as claimed in one of the preceding claims, characterized in that the fields are arranged in the form of a spirally applied CD data track on the analysis support. 33. The method as claimed in one of the preceding claims, characterized in that one or more writable data tracks are applied to the analysis support. 34. An analysis support, characterized in that it has at least one of the features described in the preceding claims. 35. A device for reading the analysis support described in claim 34, characterized by the following features: instruments for transmitted-light and/or incident-light detection; instruments for magnetic or electrical detection; an instrument for automatically finding the detection fields; an instrument for manually, semiautomatically and automatically feeding the analysis support through the device, together with suitable mechanical guidance; an instrument for recording analog signals; an instrument for digitizing analog signals; an instrument for time- and position-resolved synchronization of the recording of analog and/or digital signals; an instrument for error correction when reading and/or interpreting signals. 36. A kit, containing the essential substances for production of the analysis support described in claim 34. 37. A kit, containing the essential substances for carrying out one or more detections on an analysis support described in claim 34. |
<SOH> BRIEF DESCRIPTION OF THE INVENTION <EOH>The invention relates to a method for detecting and/or quantifying molecules from a sample on an analysis support formatted with a digital data code, wherein a detection reaction induces a change of the codewords, and these can be sequentially read and interpreted in the predetermined format. Surface-based detection methods have been established for many years in the biochemical laboratory. With the increasing demands of molecular biological research, the need for highly parallelized and miniaturized technologies for studying the binding in complex molecular mixtures is growing. The known methods for detecting and quantifying target molecules from a sample involve a plurality of steps, which are carried out by means of various devices which are sometimes elaborate and expensive. Microarrays are one of the possible ways of analyzing a multiplicity of biological molecules. Microarray technology, in which many different biological biomolecules such a DNA or proteins are applied, densely packed, in a predefined pattern on a substrate surface, has now become the standard method for parallel analysis of biological samples. This technology is used, for example, in the analysis of gene expression, in genetic diagnosis, in biological and pharmaceutical research and for the determination of genetically modified organisms in the food industry. In microarray technology, biochemical sensor molecules such as DNA or proteins are applied to metal, glass, membrane or plastic surfaces, especially polycarbonate supports. After contact with the applied sample, it is possible to detect the molecular interaction and usually to obtain information about the bound quantity and/or about the strength of the interaction. According to the prior art, the binding is usually detected via the generation and detection of an optical signal. In this case, it is conventional to use a microscope or a functionally similar device, especially a CD reader head (WO00/26677, WO00/36398). The information about binding which has or has not taken place at a particular site is obtained by image processing, which is typically carried out by computer software after analog-digital conversion in the case of microarrays with a relatively high density. In one of the known methods, the information about binding which has or has not taken place at a particular site is labeled by the accumulation of grains (beads) at the site of the reaction of the analyte with the carrier-bound sensor molecule (for example EP918885 and Taton, Mirkin, Letsinger, Science 289: 1757-1760 (2000)). According to the prior art, beads bound in such a way are either identified directly as bodies or cause a chemical reaction such as a color change or a dyestuff precipitate. These are essentially detected by photometric methods. A camera and microscope combination is used to generate data, which can be evaluated by image analysis in the computer. One of the disadvantages of the known methods for evaluating microarray analyses is the use of complicated and expensive devices and software for detecting and evaluating very weak signals, for example emission by a few molecules of fluorescent dyestuffs. These readers and evaluation devices are the result of many years of development work, use elaborate methods such as confocal scanning microscopy, and are very expensive. These devices furthermore require special software for identifying and localizing molecule spots and for interpreting and integrating detected signals. In the next few years, initial results from gene research will start to be used in medical diagnosis and prognosis. Above all, simple and fast molecular detection methods will be required for various clinical problems. There is therefore a need for a simple, inexpensive and at least partly automated method for detecting and analyzing molecules in complex mixtures. It is an object of the present invention to provide a fast and simple method for analyzing and detecting analytes, which offers reliable interpretation of the results according to defined criteria as well as quantitative information. In order to achieve this object, the invention relates to a method with the features mentioned in claim 1 . Refinements of the invention are the subject matter of dependent claims which, like the abstract, have been worded with reference to the content of the description. The above object is achieved according to the invention by a method in which a molecular interaction at a particular site on the support leads to the generation of detectable structures, referred here as signaling elements, which can be read and interpreted in the context of the format defined in the form of a digital data code on the analysis support. Detection fields with the sensor elements needed for the respective detection, as well as other data structures, are applied in a digital format on the analysis support and are combined to form sequences of format structures which can be clearly interpreted as codewords. In order to detect or quantify an analyte in a sample, the latter is applied to the analysis support and modulation of the digital signal level on the respective detection fields is initiated with the aid of a signaling element. In this way, one codeword is replaced by another codeword allowed within the set of valid codewords. The information about a reaction which has or has not taken place is based on comparison of the respective format structures before and after the detection. The exemplary embodiments are a CD, a magnetic card, an optical card and a barcode card. The method according to the invention has the advantage, over known methods, that elaborate two-dimensional image analysis since digital data are obtained. The provision of a digital information structure obviates analog signal processing, which is complicated and prone to errors. Reliable interpretation of the results according to defined criteria is facilitated. The analysis support and the reader, which is constructed from standard components in the consumer goods industry, can furthermore be adapted to one another easily as a function of the problem, so that various tests can be developed and produced quickly and inexpensively in mass production. Other advantages, features and possible applications of the invention will be described below with the aid of the detailed description and exemplary embodiments, with reference to the drawings. In the drawings: FIG. 1 : A) shows a schematic representation of an exemplary sequence of format elements on an analysis support, which consists of blank fields 5 , address fields 6 (gray squares) and detection fields 7 (white squares), and which forms a track. The header region 8 is used for finding and initializing the track when reading; B) before detection, the sequence of format elements represents the codeword A; C) after detection has taken place, the represented sequence of format elements may represent the word A, if no reaction takes place (I.), or the codeword B in the event of a reaction (II.). The binary codewords A and B differ by a change of the signal level at the fourth position, which corresponds to a detection field. FIG. 2 : A) shows a schematic representation of the simplest embodiment of the analysis support 1 , with a track 2 of format elements, sketched linearly by way of example, and a header region 8 ; B) shows a schematic representation of another possible configuration of the analysis support 1 with a track 2 of format elements and a header region 8 , an optional central hole 3 for combination of the analysis support and a CD, together with a spiral data track 4 for the data storage and software; C) shows a schematic representation of an example of another preferred embodiment of the analysis support 1 with a central hole 3 , a spiral data track 4 in the CD-R standard for the data storage and software, and a spiral track 32 of format elements in the CD-R standard; D) shows a schematic representation of another possible configuration of the analysis support in the form of a CD 33 with a central hole 3 , a spiral data track 4 in the CD-R standard for the data storage and software, and a spiral track 32 of format elements in the CD standard. FIG. 3 : shows a schematic representation of an analysis support 1 with a plurality of tracks arranged in parallel, which consist of rows of detection, information and blank fields. Exemplary filling of the analysis support 1 by means of a microfluidic plate 9 with sensor elements or analyte solutions through microchannels 10 embedded in the support, or otherwise spatially arranged 10 , is represented. FIG. 4 : shows a schematic representation of a detection according to Example 1 . A binding reaction between a sensor element 34 , here an antibody, applied to the analysis support 1 and an analyte molecule 12 , here a protein, from the sample is represented. The detection is carried out in a sandwich immunoassay with the aid of a second antibody 11 , which is directed against a different epitope of the protein. A colloidal gold particle 13 coupled to the second antibody leads to the deposition of a silver grain 14 , which is used as a signaling element. FIG. 5 : shows a schematic representation of a detection according to Example 3 . A binding reaction between a receptor 15 applied to the support 1 and a ligand 16 from the sample, which is coupled to a hapten 17 , here biotin, is represented. After addition of a mixture of streptavidin 18 and biotinylated ferretin 19 , signaling elements are produced in the form of molecular complexes 20 . FIG. 6 : shows signaling elements for the example of polystyrene beads with a diameter of 1 μm, which are detected according to two different methods and interpreted in binary form. A) shows an image recorded by a CD reader head; B) shows the same beads recorded by fluorescence microscopy; C) shows a three-dimensional representation of the data from A), which can be interpreted in binary form. FIG. 7 : A) shows a simplified schematic representation of an optical system for detecting the reflection signal, which represents a conventional CD reader head, consisting of a laser (L) 25 , a detector for focal adjustment and signal detection (D/F) 27 , a focusing instrument 28 and a semisilvered plate 29 . The analysis support 1 with the silvering 30 is represented in the beam path in the side view; B) shows a simplified schematic representation of an optical system for detecting the transmission signal, consisting of a CD pickup from Example A), the actual detector for the transmission measurement (D) 26 and optionally an additional focusing instrument 31 . The analysis support 1 with the silvering 30 is represented in the beam path between the focusing instruments 28 and 31 in the side view. FIG. 8 : shows a schematic representation of an analysis support, on which the detection fields are applied as parallel strips so that they can be read using a barcode reader. A) The individual detection fields 23 are arranged in relatively large detection regions the form of test strips 22 . They may be fitted in a test unit together with address fields 21 which, for example, contain information about test specifications, codings (dongle) or product identification. After a detection reaction with analytes from different samples 1 and 2 , different patterns of signaling elements are obtained on the test strips, and these can be read using a barcode reader; B) The test strips 22 may consist of a plurality of detection fields 24 , which are arranged orthogonally to the main reading direction and, for example, may contain graded concentrations. FIG. 8 : shows an image of fine lines in the micrometer range, which are attributable to an antibody-antigen reaction with subsequent silver deposition. A) shows an analog image recorded by a CD reader head; B) shows analog image lines from A), read along the horizontally dashed line; C) shows digital processing of the image line from B), produced from the analog signal by using a threshold criterion, identified by the horizontal line in B). detailed-description description="Detailed Description" end="lead"? |
Method for providing services in a data transmission network and associated components |
The invention relates to, among other things, a method according to which an access function (36) for a number of service user computers (18) permits a connection between the service user computer (18) and a service provider computer (22 to 26), which is selected by a service user (A), according to requests submitted by a service user computer (18). The insertion of an access function (36), and the use of a test unit (38) make it possible to secure useful data that is to be processed in a reliant manner. |
1. A method for providing services in a data transmission network, comprising: enabling a connection between one of several service user computers and one of several service provider computers, which can be selected by a service user depending on requests from one of the service user computers with an access function; storing secure user data for the service users being stored in a central database, the secure user data being necessary for the provision of services of service provider computers; after establishing a connection between a service user computer and a selected service provider computer, as part of the service provision for the service user using the service user computer, submitting a request that can be processed only by using the secure user data of a service user to a test unit used by several service provider computers; transmitting the result of the processing to the service provider after the request has been processed by the test unit by accessing the secure user data of the service user; and providing a service by the service provider computer depending on the processing result. 2. The method in accordance with claim 1, wherein the service provider computers belong to various operators, further comprising: checking a selected service provider's authorization to submit a request using an authorization check procedure, and transmitted transmitting the processing result only if authorization exists. 3. The method in accordance with claim 1, wherein the secure user data is stored encrypted, and the service provider computer has no access to the digital key necessary for decrypting the secure user data. 4. The method for providing services in a data transmission network according to claim 1 wherein service user data containing service-related data for the service users of individual service provider computers is stored in a database, after selection of a service provider computer its authorization to receive service user data relevant to the service it provides is checked, if authorization is present the service user data of the particular service user that selected the selected service provider computer is transmitted to the selected service provider computer, and the service provider computer provides its service by using the transmitted service user data. 5. The method in accordance with claim 4, wherein the service user data is stored and transmitted encrypted, and different service provider computers use different digital keys for decrypting service user data. 6. The method in accordance with claim 4, wherein the service user data is encrypted using a central encryption process, and for encryption in accordance with the central encryption process, the same digital key is used for the service-service user data of different service provider computers. 7. The method in accordance with claim 4, wherein digital data regarding payment procedures for different service provider computers is stored in a database used by several service provider computers. 8. The method according to claim 1, wherein, the authorization of the service user is checked by using an authorization check procedure, and the selection is allowed only if authorization is present. 9. The method in accordance with claim 1, wherein the authorization check is performed using digital keys that have been generated by at least one certification center, and the certification center is part of a certification infrastructure. 10. The method in accordance with claim 9, wherein a secret digital key is used for encryption, and the secret digital key is stored in an electronically protected storage unit. 11. The method in accordance with claim 10, wherein the protected storage unit is part of a chip card with a processor, and the protected storage unit can be accessed after an authorization check only by the processor. 12. The method in accordance with claim 1, wherein the request refers to the securing of a payment. 13. The method in accordance with claim 12, Wherein the test unit, when processing the requirement, submits a request to a certification computer to receive a payment certificate, the certifying computer generates a digital payment certificate that secures the payment, and the payment certificate is transmitted via the test unit to the service provider computer. 14. The method in accordance with claim 12, wherein the test unit, when processing the request, generates a payment certificate that secures the payment, and the payment certificate is transmitted to the service provider computer. 15. The method in accordance with claim 13, wherein the payment certificate is generated with the aid of a digital key. 16. The method in accordance with claim 1, wherein the service provider computers perform the function of electronic sales platforms for various products or product groups and/or electronic service provision platforms for various services or service groups. 17. A computer readable medium upon which is stored a program with an instruction sequence that when executed by a processor performs the method of claim 1. 18. A data processing system characterized by a program in accordance with claim 17. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Until now, it has been usual for each business to have its own access function and for the customer data of each business to be stored individually, and therefore several times under certain circumstances. The security of customer data where storage of customer data is distributed in this way can be guaranteed only to a limited extent. Because of these limitations with regard to security, a trade in customer data has developed. Such trade substantially reduces the acceptance of providing services through the Internet, particularly if customer data is used in conjunction with the purchasing power, credit limit or other financial data of the customer. |
<SOH> SUMMARY OF THE INVENTION <EOH>According to an aspect of the invention, a simpler method of providing services in a data transmission network, that particularly guarantees to protect customer data from misuse better than previous methods, is provided. Furthermore, an associated program and associated data processing system are also specified. According to a further aspect of the invention, the substantial expense required to secure customer data is considered, which would reduce acceptance of the provision of services through the Internet on the part of the service provider. But to counteract this, an access function is provided that enables a connection between a service user computer and a service provider computer that can be selected from several by a service user. Furthermore, a central database may be set up in which user data to be secured for the various service users is stored, that is necessary for the provision of the services of various service provider computers. By this centralization of the access function and the database, the cost for securing customer data can be spread over a number of different service providers. The acceptance on the part of the service provider thus increases. By using the central database, the service user can also be assured that his data is protected against misuse. This thus increases the acceptance by the service user of the method of providing a service by a data transmission network. The method in accordance with an aspect of the invention is also based on the consideration that secure customer data is in fact necessary as part of the service provision, but does not necessarily have to be provided to the service provider. Therefore, the method in accordance with an aspect of the invention of providing a connection between a service user computer and a selected service provider computer as part of the provision of a service requires a central test unit for the service user using the service user computer. This requirement, for example, includes the assurance of the ability of the service user to pay. The request can be processed only by access to the secure user data of the service user. Thus, for example, cover notes from a bank are to be stored for subsequent verification purposes. On the other hand, an earlier cover note can also be read provided it is still valid. A test unit that works independently of the service provider computers processes the requirement by access to the secure user data of the service user. Only the result of the processing, but not the secure user data itself, is communicated by the test unit to the service provider computer making the request. The relevant service provider computer then provides its service depending on the result of the processing. This procedure therefore means that the secure customer data itself does not have to be communicated to a service provider computer. Only the test unit has access to the secure data. Therefore, trading with the secure customer data is hindered and misuse is effectively prevented. According to a further aspect of the invention, the service provider computers belong to different operators. After a service provider computer has been selected, its authority to make requests is checked by an authorization check procedure. The result of the processing is communicated by the test unit to the service provider computer only if authorization is present. If authorization is absent, no processing result is communicated. The request must not be processed if authorization is absent. Checking the authorization on the part of the service provider computer means that it can be guaranteed that no requests are made by unauthorized persons who could then misuse the results of the processing. According to another aspect of the invention, the secure user data is stored encrypted. The service provider computers have no access to a digital key required for encryption. The encryption procedure, or a key to be used, can be kept secret by structural and/or electronic security measures. Even if the secure customer data is copied by unauthorized persons, such persons are not in possession of the key required for decryption. The secure data thus remains protected against misuse despite the unauthorized copying. According to another aspect of the invention, service user data containing service-related data for the service users of individual service provider computers is stored in a database. After a service provider computer is selected, its authorization to receive service user data relative to the service it provides is checked. The requested service user data is communicated to the selected service provider computer only where authorization exists. Only the service-related data of the particular service user that has selected the selected service provider computer is communicated. The service provider computer then provides its service by using the communicated service user data. By checking the authorization to receive service user data, it can be guaranteed that the service user data of individual service providers is not improperly communicated to third parties. According to another aspect of the invention, the database for storing the service user data is part of the central database. In yet another aspect of the invention, the same method of checking is used for checking the authorization for making requests and for checking the authorization for receiving service-related service user data. Thus, only one authorization check procedure has to be carried out in each case. In a development of the method with a database for service user data, the service user data is stored encrypted and is also transmitted encrypted. Different service provider computers use different digital keys for decrypting the service user data. This measure guarantees that the service user data can be decrypted only by the authorized service provider. Other service provider computers, and also the operator of the databases, are not able to decrypt the service user data. This thus effectively protects the service user data from misuse. The storage of the service user data outside the business providing the service is thus accepted more readily. According to a further aspect of the invention where service user data is used, the service user data is additionally or alternatively encrypted by a central encryption process. A digital key to which the service provider computer has no access is used for decrypting the user data encrypted using the central encryption process. In this way, both the unencrypted data from the service provider computers and encrypted data can be securely stored using the same central process. A double encryption also offers additional security against the misuse of service-related data. According to a further aspect of the invention, digital data regarding payment procedures for different service provider computers is stored in a database used by several service -provider computers. This database is, for example, part of the central database. The aforementioned encryption process can also be used to secure data regarding payment procedures. Furthermore, an authorization check is carried out before the data on payment procedures is transmitted. According to yet another aspect of the invention, the authorization of the service user is checked by using an authorization check procedure. The selection is permitted only if authorization is present. This authorization check prevents misuse by the service user. In another aspect of the invention, the authorization check(s) is/are carried out using digital keys that have been generated by at least one certification center. The certification center itself is part of a certification chain. Compared with using passwords, the use of digital keys offers an increased safety, and an additional safety if passwords are additionally used. A certification infrastructure can, for example, be set up in accordance with standard X.509 of the ITU-T (International Telecommunication Union—Telecommunication Sector). Other infrastructures are also used, such as an infrastructure in accordance with the specifications of the IETF (Internet Engineering Task Force) in Request for Comment 2459, January 1999. Setting up such infrastructures and including them in the method in accordance with the invention guarantees a high degree of security for all participating sides. For example, invalid keys can be easily blocked. According to still a further aspect of the invention, a secret digital key can be used for encryption. The secret key is stored in a secure electronic storage unit. In one embodiment, the secure storage unit is part of a chip card containing an embedded processor and a secure storage unit. It is possible to read from, and write to, the secure storage unit by this processor. In another embodiment, an authorization check is carried out before access, that for example contains a request for a password or secret number. Preferably, an asymmetric coding method is used. According to a further aspect of the invention, the request refers to securing a payment. Securing payment is the core of the service provision using a data transmission network and is therefore particularly important for the acceptance of this method. There is therefore a requirement that a third party accepts responsibility if the service user does not pay for the service used. With one embodiment, these guarantees are time-limited, e.g. to one day or to the duration of a connection between the service user and service provider computer. According to a further aspect of the invention, the test unit requests receipt of a payment certificate to a certification computer when processing the request. The certification computer generates a digital payment certificate that guarantees the payment. The payment certificate is then passed on through the test unit to the service provider computer. In one embodiment, encryption and/or signature methods using digital keys are also used to generate the digital payment certificate. Also, in one embodiment, the certification computer is part of a certification infrastructure. The certificates generated by the certification computer have a shorter period of validity than the certificates for the digital keys. Misuse of the payment certificate or payment attribute is better prevented by the short period of validity. A certification computer in one embodiment is a TrustedA (Trusted Authorizer) computer, such as is sold by the Irish company SSE, see www.sse.ie. According to an alternative aspect of the invention, the test unit itself generates a payment certificate that guarantees payment when processing the request. In this case, the test unit is, for example the property of a banking institute or credit institute. The payment certificate generated by the test unit is also passed on to the service provider computer. The service provider computer then, for example, checks the payment certificate and initiates the provision of the service, provided the payment certificate is valid and the request is confirmed. In another aspect of the invention, the service providers perform the functions of electronic sales platforms and/or electronic service platforms, e.g. calling up music data, video data or program data, e-business, banking transactions, commercial transactions, information services secure digital voice transmission. In this way, the access function offers the service user access, for example to a virtual shopping mall. The method in accordance with the invention is, however, also used for other services where secure data of the service user is part of the service provision, for example credit businesses. The invention also relates to a program with a sequence of instructions, the execution of which by a processor is performed by the method in accordance with the different aspects of the invention. Furthermore, a data processing system containing such a program is protected. The aforementioned technical actions therefore apply for the program and the data processing system. Asymmetric methods of encryption, e.g. the RSA method (Revist, Shamir, Adleman) can be used for encryption. Symmetric methods, such as the triple DES (Data Encryption Standard) algorithm can also be used. Another common encryption method is, for example, the ECC (Elliptic Curve Cryptography) method. |
Method of building a tyre and tyre for a two-wheeled vehicle |
A method of building a tyre includes forming a carcass structure comprising at least one carcass ply, applying a belt structure at a radially external position with respect to the at least one carcass ply, applying a pair of sidewalls at an axially external position with respect to side surfaces of the at least one carcass ply, applying a tread band to the belt structure at a radially external position of the belt structure, and applying annular stiffening inserts against the side surfaces of the at least one carcass ply before applying the sidewalls. Ends of the at least one carcass ply engage respective circumferentially inextensible annular anchoring structures. Each sidewall extends radially away from one of the annular anchoring structures. Each annular insert extends between one of the annular anchoring structures and a corresponding edge of the belt structure. A tyre for a two-wheeled vehicle is also disclosed. |
1-30. (canceled) 31. A method of building a tyre, comprising: forming a carcass structure comprising at least one carcass ply; applying a belt structure at a radially external position with respect to the at least one carcass ply; applying a pair of sidewalls at an axially external position with respect to side surfaces of the at least one carcass ply; applying a tread band to the belt structure at a radially external position of the belt structure; and applying annular stiffening inserts against the side surfaces of the at least one carcass ply before applying the sidewalls; wherein ends of the at least one carcass ply engage respective circumferentially inextensible annular anchoring structures, wherein each sidewall extends radially away from one of the annular anchoring structures, and wherein each annular insert extends between one of the annular anchoring structures and a corresponding edge of the belt structure. 32. The method of claim 31, wherein each annular insert is formed by winding at least one continuous thread element into concentric coils. 33. The method of claim 31, wherein the annular inserts are made together with at least one belt layer of the belt structure by winding at least one continuous thread element into coils distributed between the annular anchoring structures. 34. The method of claim 31, wherein the annular inserts are applied before the belt structure is applied. 35. The method of claim 34, wherein during applying the belt structure, the edges of the belt structure are each superposed on a radially external edge of one of the annular inserts. 36. The method of claim 32, wherein the at least one thread element is wound with a substantially constant pitch, so that the coils are substantially spaced apart a same distance from each other. 37. The method of claim 32, wherein the at least one thread element is wound with a pitch that has a lower value close to the annular anchoring structures than close to the edges of the belt structure. 38. The method of claim 32, wherein the at least one thread element is wound with a pitch that has a higher value close to the annular anchoring structures than close to the edges of the belt structure. 39. The method of claim 32, wherein the at least one thread element is wound with a varying pitch between the annular anchoring structures and the edges of the belt structure. 40. The method of claim 31, wherein the annular inserts constitute part of the belt structure, and wherein the annular inserts extend continuously between the annular anchoring structures. 41. The method of claim 40, wherein the belt structure comprises spiraled coils of cord extending from a first annular anchoring structure to an axially opposite second annular anchoring structure. 42. The method of claim 40, wherein the belt structure consists of spiraled coils of cord extending from a first annular anchoring structure to an axially opposite second annular anchoring structure. 43. The method of claim 31, wherein applying the annular inserts is preceded by forming an intermediate elastomer substrate against an outer surface of the at least one carcass ply. 44. The method of claim 43, wherein the elastomer substrate is formed by winding at least one continuous elongated element of elastomer material into coils disposed in mutual side-by-side, superposed, or side-by-side and superposed relationship against the outer surface of the at least one carcass ply. 45. The method of claim 31, wherein forming the carcass structure comprises: preparing strip lengths; disposing the strip lengths, circumferentially distributed, on a toroidal support to form the at least one carcass ply; and applying the annular anchoring structures close to inner circumferential edges of the at least one carcass ply; wherein each strip length comprises longitudinal and parallel thread elements at least partly coated with at least one layer of elastomer material, and wherein each of the strip lengths extends in a substantially U-shaped configuration around a cross-section outline of the toroidal support to define two side portions, mutually spaced apart in an axial direction, and a crown portion, extending at a radially external position between the side portions. 46. The method of claim 45, wherein forming the at least one carcass ply comprises: disposing a first series of the strip lengths on the toroidal support; applying at least first annular anchoring inserts of the annular anchoring structures against end flaps of the strip lengths of the first series; and disposing a second series of the strip lengths on the toroidal support; wherein the first series of strip lengths are circumferentially distributed with a circumferential pitch corresponding to a multiple of a width of the strip lengths of the first series, wherein each strip length of the second series comprises end flaps that are superposed on respective first annular inserts at an axially opposite position relative to the end flaps of the strip lengths of the first series. 47. The method of claim 46, wherein the strip lengths of the first series are disposed in first deposition planes, wherein the strip lengths of the second series are disposed in second deposition planes, wherein the first deposition planes are offset in parallel from a plane radial to a rotation axis of the toroidal support, wherein the second deposition planes are offset in parallel from the plane radial to the rotation axis of the toroidal support, and wherein the first and second deposition planes are offset on opposite sides of the plane radial to the rotation axis of the toroidal support. 48. The method of claim 46, wherein the annular inserts are applied against the side portions of the strip lengths of the first series before disposing the second series of the strip lengths. 49. A tyre for a two-wheeled vehicle, comprising: a carcass structure comprising at least one carcass ply; a belt structure applied at a radially external position with respect to the at least one carcass ply; a pair of sidewalls applied at an axially external position with respect to side surfaces of the at least one carcass ply; a tread band applied to the belt structure at a radially external position of the belt structure; and annular stiffening inserts axially interposed between the side surfaces of the at least one carcass ply and the sidewalls; wherein ends of the at least one carcass ply engage respective circumferentially inextensible annular anchoring structures, wherein each sidewall extends radially away from one of the annular anchoring structures, and wherein each annular insert extends between one of the annular anchoring structures and a corresponding edge of the belt structure. 50. The tyre of claim 49, wherein each annular insert comprises at least one continuous thread element wound into concentric coils. 51. The tyre of claim 50, wherein the at least one thread element extends past the annular insert to form at least one belt layer of the belt structure. 52. The tyre of claim 49, wherein the belt structure comprises edges that are superposed on radially external edges of the annular inserts. 53. The tyre of claim 50, wherein the coils of the at least one thread element are substantially spaced apart a same distance from each other. 54. The tyre of claim 50, wherein the coils of the at least one thread element comprise a distribution pitch that has a lower value close to the annular anchoring structures than close to the edges of the belt structure. 55. The tyre of claim 50, wherein the coils of the at least one thread element comprise a distribution pitch that has a higher value close to the annular anchoring structures than close to the edges of the belt structure. 56. The tyre of claim 49, further comprising at least one intermediate elastomer substrate interposed between the at least one carcass ply and the annular inserts. 57. The tyre of claim 50, wherein the at least one thread element comprises at least one metal cord of a 3×3×0.175 HE HT type. 58. The tyre of claim 49, wherein the at least one carcass ply comprises: a plurality of strip lengths circumferentially distributed around a geometric axis of the tyre; wherein each strip length comprises longitudinal and parallel thread elements at least partly coated with at least one layer of elastomer material, and wherein each of the strip lengths extends in a substantially U-shaped configuration of a cross-section outline of the carcass structure to define two side portions, mutually spaced apart in an axial direction, and a crown portion, extending at a radially external position between the side portions. 59. The tyre of claim 58, comprising: a first series of the strip lengths; and a second series of the strip lengths; wherein each of the annular anchoring structures comprises at least one annular insert axially interposed between the strip lengths of the first series and the strip lengths of the second series. 60. The tyre of claim 59, wherein the strip lengths of the first series are disposed in first deposition planes, wherein the strip lengths of the second series are disposed in second deposition planes, wherein the first deposition planes are offset in parallel from a plane radial to the geometric rotation axis of the tyre, wherein the second deposition planes are offset in parallel from the plane radial to the geometric rotation axis of the tyre, and wherein the first and second deposition planes are offset on opposite sides of the plane radial to the geometric rotation axis of the tyre. 61. The tyre of claim 59, wherein the annular inserts are axially interposed between the side portions of the strip lengths of the first series and the side portions of the strip lengths of the second series. |
Replicons derived from positive strand rna virus genomes useful for the production of heterologous proteins |
The present invention relates to replicons or self-replicating RNA molecules, derived from the genome of cardioviruses and aphtoviruses, which can be used to express heterologous proteins in animal cells. When injected in an animal host, for example in the form of naked RNA, these replicons permit the translation of the encoded heterologous protein. If the encoded heterologous protein is a foreign antigen, these replicons induce an immune response against the encoded heterologous protein. The invention uses cardiovirus and aphtovirus genomes to construct these replicons. The invention demonstrates that these replicons, when injected as naked RNA, can induce immune responses against a replicon-encoded heterologous protein in an animal recipient without the help of any kind of carrier or adjuvant. |
1. A self-replicating recombinant positive strand RNA molecule of a viral genome of an RNA virus, wherein the RNA molecule comprises: a) RNA sequence encoding the non-structural proteins of the RNA virus; b) viral non-encoding RNA sequences necessary for viral replication; and (c) RNA sequence encoding a heterologous protein or fragment of a heterologous protein. 2. A self-replicating recombinant positive strand RNA molecule of a viral genome of an RNA virus, wherein the RNA molecule comprises: (a) RNA sequence encoding the non-structural proteins of the RNA virus; (b) viral non-encoding RNA sequences necessary for viral replication; wherein the RNA sequence in a) and/or the viral non-encoding RNA sequences in b) are either in mutated or truncated forms, and (c) RNA sequence encoding a heterologous protein or fragment of a heterologous protein. 3. The self-replicating recombinant positive strand RNA molecule according to claims 1 or 2, wherein the RNA virus is in the genus of Cardiovirus or Aphtovirus. 4. The self-replicating recombinant positive strand RNA molecule of claim 3, wherein the RNA virus is a Mengo virus. 5. The self-replicating recombinant positive strand RNA molecule of claim 4 further comprising the Cis-acting Replication Element (CRE) of the Mengo virus VP2 gene. 6. The self-replicating recombinant positive strand RNA molecule of claim 4 further comprising the Cis-acting Replication Element (CRE) of the Theiler's virus VP2 gene. 7. The self-replicating recombinant positive strand RNA molecule according to claims 1 or 2, wherein the heterologous protein is chosen from a biologically active protein, a reporter protein, a cytotoxic protein, a protein of a pathogen, or a protein of a tumor. 8. The self-replicating recombinant positive strand RNA molecule of claim 7, wherein the reporter protein is green fluorescent protein. 9. The self-replicating recombinant positive strand RNA molecule of claim 7, wherein the protein of a pathogen is influenza nucleoprotein or influenza hemagglutinin. 10. The self-replicating recombinant positive strand RNA molecule according to claims 1 or 2, wherein the heterologous protein fragment is an antigen or epitope of said heterologous protein. 11. A vaccine comprising at least one self-replicating recombinant positive strand RNA molecule according to claims 1 or 2, and a pharmaceutically acceptable carrier. 12. The vaccine of claim 11, wherein the self-replicating recombinant positive strand RNA molecule is naked RNA. 13. The vaccine of claim 11, wherein the self-replicating recombinant positive strand RNA molecule is encapsidated. 14. The vaccine according to claims 11, wherein the pharmaceutically acceptable carrier is chosen from water, petroleum oil, animal oil, vegetable oil, peanut oil, soybean oil, mineral oil, sesame oil, saline solutions, aqueous dextrose, glycerol solutions, polycationic particles, protein particles, protamine particles, liposomes, and gold particles. 15. A method of inducing a protective immune response in a host comprising: (a) at least one self-replicating recombinant positive strand RNA molecule of claims 1 or 2 in a pharmaceutically acceptable carrier, and b) immunizing the host with the preparation of step (a). 16. A method of inducing an immune response in a host according to claim 15, wherein the self-replicating recombinant positive strand RNA molecule of step (a) is prepared in naked form. 17. A method of inducing an immune response in an a host according to claim 15, wherein the self-replicating recombinant positive strand RNA molecule of step (a) is an encapsidated RNA. 18. The method according to claim 15, wherein the pharmaceutically acceptable carrier is chosen from water, petroleum oil, animal oil, vegetable oil, peanut oil, soybean oil, mineral oil, sesame oil, saline solutions, aqueous dextrose, glycerol solutions, polycationic particles, protein particles, protamine particles, liposomes, and gold particles. 19. The method according to claim 15, wherein the host is a human, a pig, a dog, a cat, a cow, a chicken, a mouse, or a horse. 20. A DNA molecule that encodes a self-replicating recombinant positive strand RNA molecule of a viral genome of an RNA virus, wherein the RNA molecule comprises: (a) RNA sequence encoding the non-structural proteins of the RNA virus; (b) viral non-encoding RNA sequences necessary for viral replication; and (c) RNA sequence encoding a heterologous protein or fragment of a heterologous protein. 21. A DNA molecule that encodes a self-replicating recombinant positive strand RNA molecule of a viral genome of an RNA virus, wherein the RNA molecule comprises: (a) RNA sequence encoding the non-structural proteins of the RNA virus; (b) viral non-encoding RNA sequences necessary for viral replication; wherein the RNA sequence in a) and/or the viral non-encoding RNA sequences in b) are either in mutated or truncated forms, and (c) RNA sequence encoding a heterologous protein or fragment of a heterologous protein. 22. The DNA molecule according to claims 20 or 21, wherein the RNA virus is in the genus of Cardiovirus or Aphtovirus. 23. The DNA molecule according to claim 22, wherein the RNA virus is a Mengo virus. 24. The DNA molecule of claim 23, further encoding RNA comprising the Cis-acting Replication Element (CRE) of the Mengo virus VP2 gene. 25. The DNA molecule of claim 23, further encoding RNA comprising the Cis-acting Replication Element (CRE) of the Theiler's virus VP2 gene. 26. The DNA molecule according to claims 20 or 21, wherein the heterologous protein is chosen from a biologically active protein, a reporter protein, a cytotoxic protein, a protein of a pathogen, or a protein of a tumor. 27. The DNA molecule of claim 26, wherein the reporter protein is green fluorescent protein. 28. The DNA molecule of claim 26, wherein the protein of a pathogen is influenza nucleoprotein or influenza hemagglutinin. 29. The DNA molecule of claim 26, wherein the heterologous protein fragment is an antigen or epitope of said heterologous protein. 30. The DNA molecule of claim 26, further comprising a suitable cloning vector. 31. A DNA molecule comprising the sequence of SEQ. ID. NO. 26 (deposited at CNCM, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France, on May 21, 2001, under Accession No.1-2668) or a fragment thereof, and DNA sequence encoding a heterologous protein or fragment of a heterologous protein in an expressible form. 32. A DNA molecule comprising the sequence of SEQ. ID. NO. 26 (deposited at CNCM, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France, on May 21, 2001, under Accession No. I-2668) either in a mutated or truncated form or a fragment thereof and DNA sequence encoding a heterologous protein or fragment of a heterologous protein in an expressible form. 33. The DNA molecule according to claims 31 or 32, wherein the heterologous protein is chosen from a biologically active protein, a reporter protein, a cytotoxic protein, a protein of a pathogen, or a protein of a tumor. 34. The DNA molecule according to claim 33, wherein the reporter protein is green fluorescent protein. 35. The DNA molecule according to claim 33, wherein the protein of a pathogen is influenza nucleoprotein or influenza hemagglutinin. 36. The DNA molecule according to claims 31 or 32, wherein the heterologous protein fragment is an antigen or epitope of said heterologous protein. 37. A DNA molecule comprising the sequence of SEQ. ID. NO. 27 (deposited at the CNCM, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France, on May 21, 2001, under Accession No. I-2669) or a fragment thereof and DNA sequence encoding a heterologous protein or fragment of a heterologous protein. 38. A DNA molecule comprising the sequence of SEQ. ID. NO. 27 (deposited at the CNCM, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France, on May 21, 2001, under Accession No. I-2669) either in a mutated or truncated form or a fragment thereof and DNA sequence encoding a heterologous protein or fragment of a heterologous protein in an expressible form. 39. The DNA molecule according to claims 37 or 38, wherein the heterologous protein is chosen from a biologically active protein, a reporter protein, a cytotoxic protein, a protein of a pathogen or a protein of a tumor. 40. The DNA molecule according to claim 39, wherein the protein of a pathogen is influenza nucleoprotein or influenza hemagglutinin. 41. The DNA molecule according to claims 37 or 38, wherein the heterologous protein fragment is an antigen or epitope of said heterologous protein. 42. A method of inducing a protective immune response in a host comprising: (a) providing a preparation comprising at least one DNA molecule of claims 20 or 21 in a pharmaceutically acceptable carrier; and (b) immunizing the host with the preparation of step (a). 43. A method of inducing a protective immune response in a host according to claim 42, wherein the DNA molecule is naked DNA. 44. A method of inducing a protective immune response in a host according to claim 42, wherein the DNA molecule is encapsidated. 45. A therapeutic composition comprising at least a DNA molecule according to claims 20 or 21 or a self-replicating recombinant positive strand RNA molecule according to claims 1 or 2 in an acceptable medium. 46. A therapeutic kit comprising at least a DNA molecule according to claims 20 or 21 or a self-replicating recombinant positive strand RNA molecule according to claims 1 or 2 in an acceptable medium. 47. A method for modulating the immune response in a host comprising: (a) preparing at least one molecule selected from the DNA molecule of any of claims 20 or 21 and the self-replicating recombinant positive strand RNA molecule of any of claims 1 or 2 and in a pharmaceutically acceptable carrier; and (b) immunizing the host with the preparation of step (a). 48. The method of claim 47, wherein the pharmaceutically acceptable carrier is chosen from water, petroleum oil, animal oil, vegetable oil, peanut oil, soybean oil, mineral oil, sesame oil, saline solutions, aqueous dextrose, glycerol solutions, polycationic particles, protein particles, protamine particles, liposomes, and gold particles. 49. The method of claim 47, wherein the host is a human, a pig, a dog, a cat, a cow, a chicken, a mouse, or a horse. 50. A method for improving the immunogenicity of a self-replicating recombinant positive strand RNA molecule of a viral genome of an RNA virus by producing an encapsidated self-replicating recombinant positive strand RNA molecule of a viral genome of an RNA virus comprising: (a) transfecting the self-replicating recombinant positive strand RNA molecule of any of claims 1 or 2 and or the DNA molecule of any of claims 20 or 21 into cells expressing the P1 precursor of capsid proteins; (b) preparing the encapsidated self-replicating recombinant positive strand RNA molecule from the transfected cells; and (c) immunizing a host with the preparation of step (b). 51. A method for improving the immunogenicity of a self-replicating recombinant positive strand RNA molecule of a viral genome of an RNA virus comprising: (a) condensing the self-replicating recombinant positive strand RNA molecule of any of any of claims 1 or 2; and (b) immunizing a host with the condensed RNA molecule of step (a). 52. A DNA molecule, comprising the sequence of SEQ. ID. NO. 28 (deposited at CNCM, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France, on May 16, 2002, under Accession No. I-2879). 53. The DNA molecule according to claims 36, wherein the epitope of said heterologous protein is the NP118-126 epitope of the lymphocytic choriomeningitis virus nucleoprotein. 54. The DNA molecule according to claim 41, wherein the epitope of said heterologous protein is the NP118-126 epitope of the lymphocytic choriomeningitis virus nucleoprotein. |
<SOH> BRIEF DESCRIPTION OF DRAWINGS <EOH>FIG. 1 is a schematic representation of plasmids encoding subgenomic recombinant replicons derived from the Mengo virus genome. Green fluorescent protein (GFP), HA, and NP genes are shown as hatched boxes. The CRE is shown as a stippled box. The HA protein signal peptide (SP) and HA transmembrane region (TM) are indicated by black bands. FIG. 2 is an SDS-PAGE analysis demonstrating the in vitro translation and processing of the recombinant Mengo virus polyproteins in rabbit reticulocyte lysates. Positions of molecular mass markers are indicated on the right. Mengo virus protein precursors as well as some of their major cleavage products are indicated on the left. The GFP-NP and GFP polypeptides and the influenza NP encoded by the recombinant replicons are indicated by solid arrows. FIG. 3 is a slot blot demonstrating the replication of subgenomic Mengo virus-derived replicons. At the indicated times post-transfection, cytoplasmic RNA was harvested for analysis. FIG. 4 is a fluorocytometer reading of GFP expression in HeLa cells transfected with recombinant replicon rMΔBB, rMΔBB-GFP or rMΔXBB-GFP. FIG. 5 is an SDS-PAGE analysis of an immunoprecipitated influenza NP protein expressed in [ 35 ] methionine labeled HeLa cells transfected with recombinant replicon rMΔBB-NP. Loaded samples are as follows: mock transfected HeLa cells (lane 1 ); HeLa cells transfected with replicons rMΔBB (lane 2 ). rMΔBB-NP (lane 3 ) or rMΔBB-GFP-NP (lane 4 ) and harvested at 10 hours post-transfection; mock infected HeLa cells (lane 5 ) and HeLa cells infected with A/PR/8/34 virus (lane 6 ) and harvested at 20 hours post-infection. Molecular masses and positions of the viral HA protein, the viral NP protein, and the viral M1 protein are shown on the right. FIG. 6 is a CTL assay demonstrating the induction of NP-specific CTL activity in C57BL/6 mice immunized with rMΔBB-NP. Groups of four C57BL/6 mice were immunized at three week intervals with the following vaccination protocols: 1 injection of 50 μg of pCI (◯) or pCI-NP (●) DNA; 2 injections of 25 μg of rMΔBB (□) or rMΔBB-NP (▪) RNA. Splenocytes were harvested three weeks after the last injection, stimulated in vitro and then tested for cytolytic activity in a chromium release assay against syngenic EL4 target cells loaded with NP366 peptide (a) or not (b). The percentage of specific lysis is shown at various effector: target ratios. Data shown is from one out of two experiments. Three weeks after the last injection, the frequency of influenza virus-specific CDS+ T cells was measured by the IFNγ ELISPOT assay in the presence of the immunodominant NP366 peptide (c), as described in Materials and Methods. Data are expressed as the number of SFC per 10 5 spleen cells. FIG. 7 is an ELISA demonstrating the induction of NP-specific antibodies in C57BL/6 mice immunized with rMΔBB-NP, according to the same vaccination protocol as in FIG. 6 . Titers are represented as the reciprocal of the highest dilution of pooled serum, for a given group of five or six mice, giving an optical density value at 450 nm equal to two times that of background levels in a direct ELISA test using purified split A/PR/8/34 virions as antigen. FIG. 8 is a graphical representation of the pulmonary viral loads in mice immunized with rMBBΔ-NP and then challenged with influenza virus. Open circles represent mean values of each group, bars indicate standard deviations. Data shown is from one out of two experiments. FIG. 9A is an SDS-PAGE analysis demonstrating the in vitro translation of the native form of HA in rabbit reticulocyte lysates. The influenza HA polypeptide encoded by the rMΔFM-HA recombinant replicon is indicated by a solid arrow and a non-cleaved precursor by an open arrow. FIG. 9B is a slot blot demonstrating that monocistronic Mengo virus replicons cannot express foreign glycosylated protein in transfected eukaryotic cells. At the indicated times post-transfection, cytoplasmic RNA was harvested and slot blotted onto a nylon membrane for analysis. FIG. 10 is an SDS-PAGE analysis of immunoprecipitated GFP fusion polypeptides expressed in [ 35 S] methionine labeled HeLa cells transfected with recombinant Mengo virus replicons. Loaded samples were as follows: mock-transfected HeLa cells or HeLa cells transfected with replicon RNAs rMΔBB-GFP, rMΔBB-GFP-NP118 (2 clones) or rMΔB-GFP-1cmvNP. Molecular masses (kDa) are shown on the left. FIG. 11 is an ELISPOT assay demonstrating the induction of LCMV-specific T cells in BALB/c mice immunized with rMΔBB-GFP-NP118 and rMΔBB-GFP-1cmvNP replicon RNA and, as controls, with pCMV-NP and pCMV-MG34 plasmid DNA. Three weeks after the last injection, the frequency of LCMV-specific CD8+ T cells was measured by the IFNγ ELISPOT assay in the presence of the immunodominant NP118-126 peptide, as described in Materials and Methods. Data are expressed as the number of SFC per 10 5 spleen cells. FIG. 12 is a fluorocytometer reading of GFP expression in HeLa cells transfected with recombinant Mengo virus replicons rMΔBB-GFP, rMΔBB-GFP-NP118, or rMΔBB-GFP-1cmvNP. detailed-description description="Detailed Description" end="lead"? |
Vectors for enhanced expression of Vegf for disease treatment |
The invention provides a vector which is capable of the expression of a vascular endothelial growth factor wherein said vector comprises a modified PCMV promoter. The invention further provides use of a vector which is capable of and expressing a vascular endothelial growth factor (VEGF) for the regulation of endothelial function, angiogenesis and arteriogenesis. The invention further comprises use of a vector which is capable of the expression of a vascular endothelial growth factor (VEGF) for the prophylactic treatment of arterial diseases and/or bone marrow diseases and/or neural diseases. |
1. A vector which is capable of effecting the expression of a vascular endothelial growth factor (VEGF or a functional equivalent thereof) in an appropriate environment wherein said vector comprises a modified pCMV promoter. 2. A vector according to claim 1 wherein said modification comprises the insertion of a 9 bp nucleic acid sequence ACGCGCGCT, wherein A is a deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl and T is thymidyl. 3. A vector according to claim 1 or 2 wherein said vector comprises an antibiotic resistance gene. 4. A vector according to claim 3 wherein said antibiotic resistance gene comprises kanamycin. 5. A vector according to anyone of claims 1-4 wherein said vector comprises a 5′ HSV translation initiation signal and/or a β-globin splicing/adenylation signal. 6. A vector according to anyone of claims 1-5 wherein said vector comprises a ColE1-like replicon and/or an F1 replication origin. 7. A vector according to anyone of claims 1-6 wherein said vascular endothelial growth factor (VEGF) is derived from a mammal. 8. A vector according to claim 7 wherein said mammal is a human. 9. A vector according to anyone of claims 1-8 wherein said vector comprises a vascular endothelial growth factor (VEGF) nucleic acid. 10. A vector according to anyone of claims 1-9 wherein said nucleic acid is operative in mammalian systems. 11. A vector according to claim 10 wherein said vector can replicate to a high copy number in a bacterial cell. 12. A host cell which comprises a vector according to anyone of claims 1-11. 13. A host cell according to claim 12 which is human. 14. Use of a vector according to anyone of claims 1-11 in the preparation of a medicament for the regulation of endothelial function. 15. Use of a vector according to anyone of claims 1-11 in the preparation of a medicament for the regulation of angiogenesis. 16. Use of a vector according to anyone of claims 1-11 in the preparation of a medicament for the regulation of arteriogenesis. 17. Use of a vector according to anyone of claims 1-11 in the preparation of a medicament to induce the release of hematopoietic growth factors by bone marrow endothelial cells. 18. Use of a vector according to anyone of claims 1-11 in the preparation of a medicament to induce the development of supplemental collateral blood vessels. 19. Use of a vector according to anyone of claims 1-11 in the preparation of a medicament to induce therapeutic angiogenesis 20. Use of a vector according to anyone of claims 1-11 in the preparation of a medicament to induce transendothelial progenitor cell migration. 21. Use of a vector according to anyone of claims 1-11 in the preparation of a medicament to increase endothelial fenestration. 22. A composition comprising a vector according to anyone of claims 1-11. 23. A composition according to claim 22 which is a pharmaceutical composition. 24. Use of a composition according to claim 22 or 23 and/or a vector according to anyone of claim 1-11 for the treatment of arterial diseases and/or bone marrow diseases and/or neural diseases and/or inflammatory disorders, in particular tissue ischemia especially associated with diabetes. 25. A method of treatment of arterial diseases and/or bone marrow diseases and/or neural diseases and/or inflammatory disorders, comprising administering a vector according to anyone of claims 1-11 and/or a composition according to claim 22 or 23 with a carrier to a suitable recipient. |
System and method for unattended delivery |
An inventive system and method for unattended delivery of goods includes an electronic tag associated with the item and having a first transceiver, and a drop box located at the destination for the item, the drop box having a second transceiver which wirelessly communicates with the first transceiver to allow access to the drop box. |
1. A system for unattended delivery of an item comprising: an electronic tag associated with said item and comprising a first transceiver; and a drop box located at a destination for said item, said drop box comprising a second transceiver which wirelessly communicates with said first transceiver to allow access to said drop box. 2. The system according to claim 1, further comprising: a dock to serve as a base for said drop box. 3. The system according to claim 2, wherein at least one of said dock and said drop box comprises a lock mechanism for securing said drop box to said dock. 4. The system according to claim 2, wherein said dock comprises a temperature control unit for controlling a temperature inside said drop box, and wherein said drop box comprises an insulated drop box and a port for connecting to said temperature control unit. 5. The system according to claim 1, wherein said drop box further comprises a first memory device for storing a first identification number, and wherein said electronic tag further comprises a second memory device for storing a second identification number. 6. The system according to claim 1, wherein said drop box further comprises a processor for comparing said first identification number and said second identification number, and wherein said drop box unlocks when said first identification number matches said second identification number. 7. The system according to claim 1, wherein said first and second transceivers each comprise a two-way communication analog chip. 8. The system according to claim 1, wherein said electronic tag further comprises an activating device to activate a function of said electronic tag. 9. The system according to claim 8, wherein said activating device is engaged in order to transmit data from said electronic tag to said drop box. 10. The system according to claim 1, wherein said drop box further comprises an activating device to activate a function of said drop box. 11. The system according to claim 10, wherein said activating device is engaged in order to transmit data from said drop box to said electronic tag. 12. The system according to claim 5, wherein said first and second memory devices store delivery data comprising a delivery date and delivery time. 13. The system according to claim 1, wherein said electronic tag is affixed to an outside portion of said item. 14. The system according to claim 1, further comprising: a transport vehicle for delivering said item to said destination. 15. The system according to claim 14, wherein said a transport vehicle is directed to said drop box using electronic positioning system. 16. The system according to claim 15, wherein said electronic positioning system comprises a global positioning system, and wherein said transport vehicle comprises a global positioning system receiver. 17. The system according to claim 1, wherein said first transceiver wirelessly communicates with said second transceiver via a radio frequency link. 18. The system according to claim 1, further comprising: a container for containing said goods, wherein said electronic tag is affixed to said container. 19. The system according to claim 1, further comprising: an access card comprising a third transceiver, for wirelessly communicating with said second transceiver to access said drop box. 20. The system according to claim 1, wherein said drop box further comprises a signaling device and wherein said signaling device is activated when said item arrives at a destination. 21. The system according to claim 14, wherein said transport vehicle comprises a computer system which determines an optimum route for delivering said item. 22. The system according to claim 14, further comprising: a base station comprising a fourth transceiver for wirelessly communicating with said first transceiver while said item is on said transport vehicle. 23. The system according to claim 22, wherein said transport vehicle comprises a loop antenna, and wherein said base station wirelessly communicates with said electronic tag using said loop antenna. 24. An insulated drop box for a delivery system, comprising: an insulated housing for receiving an item of goods; and a transceiver for wirelessly communicating with an electronic tag in order to access said drop box. 25. A method for unattended delivery of an item of goods comprising: associating said item with an electronic tag comprising a first transceiver; transporting said item to a destination; and placing said item in a drop box located at said destination, said drop box comprising a second transceiver which wirelessly communicates with said first transceiver to allow access to said drop box. 26. A programmable storage medium tangibly embodying a program of machine-readable instructions executable by a digital processing apparatus to perform a method for unattended delivery of goods, said method comprising: associating said item with an electronic tag comprising a first transceiver; transporting said item to a destination; and placing said item in a drop box located at said destination, said drop box comprising a second transceiver which wirelessly communicates with said first transceiver to allow access to said drop box. |
<SOH> BACKGROUND OF THE INVENTION <EOH>1. Field of the Invention The present invention relates to a system and method for delivering packages, and in particular, a system and method for delivering packages to a destination which is unattended. 2. Description of the Related Art A number of companies have developed businesses where customers select and purchase grocery items or dry goods online using the world wide web. The companies inventory these items in a local warehouse and deliver the groceries and dry goods items directly to a customer's door. Often the customer can arrange for the goods to be delivered to his home or business at a time when the customer knows he will be in. However, in some instances no such arrangement can be made. In those cases, where the destination is unattended, the goods may be, for example, delivered to a refrigerator stored in the customer's garage or outbuilding, the door to which the driver can open using a keypad entry code provided by the customer. In any event, the customer is charged a delivery fee that represents only a portion of the actual delivery costs incurred by the vendor. The vendor derives the remaining costs from margin or profit on the product itself. The rationale is that it would cost much less to operate a warehouse and cover a portion of the delivery cost from product profit margin than to run a conventional retail store and the necessary warehouses to supply the retail store. Moreover, a consumer obtains enhanced convenience over conventional retail stores since the goods are delivered directly to the customer's front door and pricing may be at least competitive with a retail store. However, delivery service companies are finding it hard to make a profit. One reason is that the cost of attended delivery is high, requiring both a high minimum order and a high delivery fee to insure profitability of the company. Unattended delivery is more popular with customers, however, delivery companies have been unable to profitably implement unattended delivery systems. Therefore, although demand for such delivery service appears to be high, few internet-based delivery services have been successful and most appear to be failing. Indeed, while some companies providing such services are beginning to claim “operational profitability”, none have actually achieved profitability and, in fact, several of the companies have even gone bankrupt. |
<SOH> SUMMARY OF THE INVENTION <EOH>In view of the foregoing and other problems, disadvantages, and drawbacks of the conventional methods and structures, an object of the present invention is to provide a system and method for unattended delivery. The present invention includes an inventive system for unattended delivery of an item. The inventive system includes an electronic tag associated with the item and having a first transceiver, and a drop box located at the destination, the drop box having a second transceiver which wirelessly communicates with the first transceiver to allow access to the drop box. The drop box may include an insulated drop box and a port for connecting to the temperature control unit. The first and second transceivers may include, for example, two-way communication analog chips, and may wirelessly communicate via a radio frequency link. The inventive system may also include a transport vehicle for transporting the item to its destination. The inventive system may also include a dock to serve as a base for the drop box. Further, the dock and/or the drop box may include a lock mechanism for securing the drop box to the dock. The dock may also include a temperature control unit for controlling a temperature inside the drop box. The drop box may also include a first memory device for storing a first identification number, and the electronic tag may include a second memory device for storing a second identification number. For example, the first and second memory devices may store delivery data such as a delivery date and delivery time. In addition, the drop box may include a processor for comparing the first identification number and the second identification number, and unlock when the first identification number matches the second identification number. The drop box may also include a signaling device (e.g., light emitting device). For example, the signaling device may be activated when the item arrives at a destination. The electronic tag may also include an activating device (e.g., switch, button, etc.) to activate a function of the electronic tag. For instance, the activating device may be engaged in order to transmit data from the electronic tag to the drop box. Likewise, the drop box may include an activating device to activate a function of the drop box. This activating device may be engaged in order to transmit data from the drop box to the electronic tag. The inventive system may also include a container for containing the item(s) of goods. In this case, the electronic tag may be affixed to the container. The electronic tag may also be affixed to an outside portion of the item. The transport vehicle may be directed to the, drop box using an electronic positioning system. For example, the electronic positioning system may include a global positioning system. In this case, the transport vehicle may include a global positioning system receiver. The inventive system may also include an access card having a third transceiver, for wirelessly communicating with the second transceiver to access the drop box. The system may also include a base station having a fourth transceiver for wirelessly communicating with the first transceiver in order to distinguish an item on the transport vehicle (e.g., by activating the signaling device on the electronic tag associated with the item). The transport vehicle may also include a computer system which determines an optimum route for delivering the item. The transport vehicle may also include a loop antenna. The base station wirelessly communicate with the electronic tag using the loop antenna. An inventive method for unattended delivery of an item of goods includes associating the item with an electronic tag having a first transceiver, transporting the item to a destination, and placing the item in a drop box located at the destination, the drop box including a second transceiver which wirelessly communicates with the first transceiver to allow access to the drop box. The present invention also includes a programmable storage medium tangibly embodying a program of machine-readable instructions executable by a digital processing apparatus to perform the inventive method for unattended delivery of goods. With its unique and novel aspects, the claimed invention provides a system and method for unattended delivery which is inexpensive to operate, resulting in lower cost to the delivery company and ultimately to consumers. |
Reactor for decomposition of ammonium dinitramide-baed liquid monopropellants and process for the decomposition |
The present invention relates to a reactor for the decomposition of ammonium dinitramide-based liquid monopropellants into hot, combustible gases for combustion in a combustion chamber, and more particularly a rocket engine or thruster comprising such reactor and a combustion chamber. The invention also relates to a process for the decompostion of ammonium dinitramide-based liquid monopropellants. |
1. A reactor for decomposition of a liquid ammonium dinitramide-based monopropellant into hot, combustible gases, comprising a hollow body (5) provided with, from the upstream end; an injector (20); a heat bed (25); and a catalyst bed (30) of porous catalyst pellets (35) which are heat resistant up to a temperature of at least 1000° C., wherein the injector is formed so as to be able to distribute the liquid propellant over the heat bed, the overall void volume is essentially formed of the porosity of the heat bed and catalyst bed, and wherein the hollow body is thermally conductive and the heat bed is in indirect thermal contact with the catalyst bed via the hollow body. 2. The reactor of claim 1, wherein the downstream portion of the catalyst bed is sintering resistant up to a higher temperature, such as up to 1200° C., more preferably up to 1400° C., and most preferably up to 1700° C. 3. The reactor of claim 1, wherein the injector has the properties of a heat bed. 4. The reactor of claim 1, wherein the pellets comprise an alumina material, more preferably, hexaaluminate, AA11O18, wherein A is an alkaline earth or rare earth metal, and preferably La. 5. Reactor of claim 4, wherein the AA11O18, is obtained by adding a solution of an aluminium alkoxide to a water-in-oil microemulsion, the aqueous phase of which comprises a solution of a water soluble salt of A, whereafter the powder formed is recovered and calcined 6. Reactor of claim 1, wherein the material comprising the pellets contains the dopant Mn. 7. Reactor of claim 1, wherein the pellets are impregnated a catalytically active metal component selected from Pt, Ru, Pd, Pt/Rh, Ir, Rh, Mn or Ir/Rh. 8. Reactor of claim 1, wherein the granules are prepared from a slurry comprising the catalyst material, a solvent, and any desired additives, by means of a drop-generating orifice to which said slurry is fed, from which orifice the drops are released by means of a relative flow of a liquid medium, and formed into spherical bodies in said liquid medium by means of the action of surface tension, and thereafter treated for consolidation. 9. A rocket engine for ammonium dinitramide-based liquid monopropellant, comprising the reactor of claim 1, wherein the overall void volume is essentially formed of the porosity of the heat bed and catalyst bed, and the combustion chamber (50). 10. The rocket engine of claim 9, wherein the porous catalytic material has been dimensionally pre-stabilised at expected operational temperature, and preferably 50° C. 11. The rocket engine of claim 9, wherein the size of the pellets is about one tenth of the inner diameter of the hollow body (5). 12. The rocket engine of claim 9, wherein the combustion chamber (50) is lined with iridium (60). 13. Rocket engine of claim 9 having a power of 0.5 to 1 kN, more preferably 0.5-50 N. 14. A process for decomposition of a liquid dinitramide-based monopropellant, comprising the steps of: (A) Subjecting the propellant to a temperature efficient for essentially bringing the propellant into the vapour phase; (B) Bringing the essentially vaporised monopropellant into contact with a porous catalytic material for decomposition of the monopropellant into hot, gaseous combustible components; and, optionally (C) Combusting the combustible components. 15. Process of claim 14, wherein heat generated from step (B) and/or (C) is used for vaporising the liquid monopropellant in step (A). 16. Process of claim 14, wherein the vaporisation of the monopropellant in step (A) is sufficient for preventing vapour induced disintegration of the porous catalytic material. 17. Process of claim 14, wherein a stabilised liquid dinitramide-based monopropellant is decomposed. |
<SOH> BACKGROUND ART <EOH>In space applications, such as rockets, satellites and other space vehicles, liquid propellant thrusters and rocket engines are often used. Such thrusters and rocket engines can for example be used for the purpose of positioning and attitude control of satellites and other space vehicles. For such purposes attitude control thrusters operating in the thrust range of typically 0.5-50 N, or Δ-V rocket engines typically operating in the thrust range of 1 N to several kN. Attitude control thrusters are required to perform short pulses or pulse trains, the duration of which typically can be fractions of seconds to several minutes. Liquid propellants can be divided into monopropellants and bipropellants. The former consists of one component, while the latter consists of two components, i.e. a liquid oxidiser and a liquid fuel. The distinction between monopropellants a bipropellants is made according to the number of components which are injected into the engine for combustion for the specific propellant. In the case of a monopropellant, which may be a mixture of several compounds or one single chemical, only one component is injected into the engine. Currently, hydrazine is about the only liquid monopropellant widely used for generation of hot gases. In the case of hydrazine the decomposition pathway occurs in two stages; first hydrazine is catalytically decomposed into hydrogen and ammonia in an exothermal reaction, and thereafter ammonia further decomposes into hydrogen and nitrogen in an endothermal reaction due to the high temperature generated in the first stage. The second stage endothermal reaction will reduce the flame temperature and reduce the specific impulse. It is therefore desirable to limit the ammonia dissociation as much as possible. When the ammonia dissociation is held to 55%, the adiabatic reaction temperature will be ca. 900° C. In a bipropellant engine, fuel and oxidizer liquids are injected, atomised and mixed in a first zone of the combustion chamber. In the case of a hypergolic bipropellant, such as hydrazine and nitrogen tetroxide, there is an initial chemical reaction in the liquid phase when a droplet of fuel impinges with a droplet of oxidiser. Bipropellants which are not hypergolic use some type of ignitor to initiate the chemical combustion. In a bipropellant system using hydrogen peroxide as the oxidiser, a catalyst may be used. Liquid bipropellants generally offer higher specific impulse than liquid monopropellants. Bipropellant systems are thus more efficient than monopropellant systems, but tend to be more complicated because of the extra hardware components needed to make sure the proper amount of fuel is mixed with the proper amount of oxidiser. Liquid monopropellants, based on a dinitramide compound, and especially ammonium dinitramide (ADN), have recently been developed, and are disclosed in WO0050363. These propellants are novel High Performance Monopropellants, which generate extremely high temperatures at proper combustion thereof. Such monopropellant comprises at least two components; a dinitramide compound (oxidiser) and a fuel. An additional solvent component may also be included, such as water. These new monopropellants, including at least two components, have been described to generate a very high temperature on combustion, such as about 1700° C. As the propellant may also include water, very high demands will be put on a suitable engine or thruster for such a fuel, consequently excluding all known monopropellant thrusters as suitable alternatives. Thus, it is an object of the present invention to provide a reactor for decomposition and combustion of liquid ammonium dinitramide-based monopropellants. It is a further object of the present invention to provide a process for decomposition ammonium dinitramide-based monopropellants, such as for rocket propulsion and for controlled gas generation for any other purpose, such as rotary power in auxiliary power units. Other objects and advantages of the present invention will become evident from the following description, examples, and the attached claims. The terms rocket engine and thruster will be used interchangeably herein to designate the portion of a liquid propellant rocket engine, in which the propellant is injected, extending downstream to the nozzle. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present inventors have investigated the decomposition pathway of ammonium dinitramide-based liquid monopropellants and found a pathway that corresponds to observed temperatures at different stages of decomposition. Consequently, it has been found that the combustion of a ammonium dinitramide-based liquid monopropellant can be divided into a series of steps, including i.a. the decomposition of the ammonium dinitramide oxidiser which eventually generates free oxygen. In a final stage, combustible components generated from thermal and catalytic decomposition will be oxidised in a homogenous gas combustion by the free oxygen thus generated. This combustion requires no catalysis. Thus, the ammonium dinitramide-based monopropellant can be regarded as being decomposed into a bipropellant, which is combusted in a final step or steps, during which the maximum temperature is reached. It has also been found that for the desired decomposition of the ammonium dinitramide oxidiser to take place, catalytic activity is required until the homogenous gas combustion. In practice, this will mean that catalytic activity will be required up to a temperature of at least 1000° C. As will be discussed in greater detail below, catalytic activity, especially in the aft portion of the catalyst, may be required may at higher temperatures, depending on the specific monopropellant. Thus, a key to this goal is the development of a suitable high-temperature catalyst. Such catalyst has now been found by the present inventors and will be described in detail below. The inventors have also developed a reactor for the decomposition of ammonium dinitramide-based liquid monopropellants, and a thruster incorporating the reactor. Such reactor and thruster will be described in detail below. The reactor could also be used for generating hot gases at high pressure for driving a turbine, vane motor, or piston motor. |
Polyolefin coated steel pipes |
Polyolefin coated steel pipes with high dynamic fracture toughness of the coating of the steel pipes during installation handling and in service, consisting of a steel pipe core, optionally an intermediate foamed plastic material, and a polyolefin coating of β-nucleated propylene copolymers whereby a test polyolefin pipe fabricated from the β-nucleated propylene copolymer has a critical pressure of >25 bars and a dynamic fracture toughness >3.5 MNm−3/2. The polyolefin coated steel pipes are suitable for off-shore transport of crude oil or gas products or district heating applications. |
1. Polyolefin coated steel pipes having high dynamic fracture toughness of the coating of the steel pipes during installation handling and in service, comprising a steel pipe core and a polyolefin coating, the polyolefin coating consisting essentially of β-nucleated propylene copolymers of from 90.0 to 99.9 wt % of propylene and 0.1 to 10.0 wt % of α-olefins of 2 or 4 to 18 carbon atoms and having melt indices of 0.1 to 8 g/10 min at 230° C./2.16 kg, a test polyolefin pipe fabricated from the β-nucleated propylene copolymer having a critical pressure of >25 bars and a dynamic fracture toughness of >3.5 MNm−3/2. 2. Polyolefin coated steel pipes according to claim 1, wherein the β-nucleated propylene copolymers comprise β-nucleated propylene block copolymers having an IRτ of the propylene homopolymer block of ≧0.98, a tensile modulus of ≧1100 MPa and a Charpy impact strength, using notched test specimens at −20° C., of ≧6 kJ/m2. 3. Polyolefin coated steel pipes according to claim 2, wherein the β-nucleated propylene block copolymers having an IRτ of the propylene homopolymer block of ≧0.98 comprise propylene copolymers obtained by polymerization with a Ziegler-Natta catalyst system comprising titanium-containing solid components, an organoalumina, magnesium or titanium compound as cocatalyst and an external donor according to the formula RxR′ySi(MeO)4-x-y, wherein R and R′ are identical or different and are branched or cyclic aliphatic or aromatic hydrocarbon residues, and y and x independently from each other are 0 or 1, provided that x+y are 1 or 2. 4. Polyolefin coated steel steel pipes according to claim 3, wherein the external donor comprises dicyclopentyldimethoxysilane. 5. Polyolefin coated steel steel pipes according to one of claims 1 to 4, wherein the β-nucleated propylene copolymers contain 0.0001 to 2.0 wt %, based on the propylene copolymers, dicarboxylic acid derivative type diamide compounds from C5-C8-cycloalkyl monoamines or C6-C12-aromatic monoamines and C5-C8-aliphatic, C5-C8-cycloaliphatic or C6-C12-aromatic dicarboxylic acids, and/or diamine derivative diamide compounds from C5-C8-cycloalkyl monocarboxylic acids or C6-C12-aromatic monocarboxylic acids and C5-C8-cycloaliphatic or C6-C12-aromatic diamines, and/or amino acid derivative diamide compounds from amidation reaction of C5-C8-alkyl-, C5-C8-cycloalkyl- or C6-C12-arylamino acids, C5-C8-alkyl-, C5-C8-cycloalkyl- or C6-C12-aromatic monocarboxylic acid chlorides and C5-C8-alkyl-, C5-C8-cycloalkyl- or C6-C12-aromatic monoamines, and/or quinacridone compounds, quinacridonequinone compounds, and/or dihydroquinacridone compounds, and/or dicarboxylic acid salts of metals from group IIa of the periodic system and/or mixtures of dicarboxylic acids and metals from group IIa of the periodic system, and/or salts of metals from group IIa of periodic system and imido acids of the formula wherein x=1 to 4; R═H, —COOH, C1-C12-alkyl, C5-C8-cycloalkyl or C6-C12-aryl, and Y═C1-C12-alkyl, C5-C8-cycloalkyl or C6-C12-aryl —substituted bivalent C6-C12-aromatic residues, as β-nucleating agent. 6. Polyolefin coated steel pipes according to claim 10, wherein the foamed plastic comprises foamed propylene copolymer having strain hardening behaviour and a melt index of 1.5 to 10 g/10 min at 230° C./2.16 kg. 7. A process for producing polyolefin coated steel pipe of claim 1 or 10, comprising preheating the steel pipe and, while rotating the preheated steel pipe melt, applying each coating onto the rotating preheated steel pipe by means of respective coating extruders each having a flat film die. 8. A process for producing polyolefin coated steel pipe according to claim 7, wherein the extruder is a cone extruder the temperature of a resultant melt of the copolymer at the ring die is from 195 to 240° C. and the temperature of the preheated steel pipe is from 160 to 200° C. 9. Installation for off-shore transport of crude oil or gas products or district heating, comprising polyolefin coated steel pipe of claim 1 or 10. 10. Polyolefin coated steel pipes of claim 1, further comprising an intermediate coating comprised of a foamed plastic. 11. A process for producing polyolefin coated steel pipe of claim 1 or 10, comprising preheating the steel pipe and applying each coating onto the preheated steel pipe from an annular die of a crosshead fed by an extruder. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Polyolefin coated steel pipes with a polyolefin coating consisting of linear low density polyethylene (JP 08,300,561), blends of propylene polymers and α-olefin copolymer elastomers (JP 2000,44,909) or syndiotactic polypropylene (JP 08,300,562) are known. The disadvantage of these polyolefin steel coatings is the insufficient dynamic fracture toughness of test pipes fabricated from the coating material. A high dynamic fracture toughness is required for coated steel pipes in order to avoid cracking of the coating during installation handling and in service. The term installation handling as used herein means any installation technique such as coiling and uncoiling of the ready made pipelines, welding and other jointing techniques and installation at the seabottom for off-shore intallations with specially designed ships, most often to a depth of several hundreds of meters, also to uncertain sea bottom conditions with risk of rock impingements etc. Installation handling of coated steel pipes, in particular for off-shore applications, involves tough conditions for the protective coating layer, including high stress, substantial elongation, surface damages, notches, impact events etc, both at low and high temperature conditions and also at high hydrostatic pressure. The coating layer is not only the layer protecting the pipeline as such from damages as mentioned, it is also doing so in a stage of high stress and/or at elevated temperatures and pressures, making the coating layer most sensitive for cracking, compare in particular the stresses induced during coiling and uncoiling. During the service life of the coated pipeline, the coating has to protect the pipeline from damages and induced stress and crack formations at conditions close to 0° C., high hydrostatic pressures where a small damage or notch in the coating could propagate into a large crack putting the pipeline as such at risk. With a high dynamic fracture toughness of the coating material the material will not crack during installation handling and in service. |
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