 bioactive state, the somatotropin, e.g., in the form of inclusion bodies, is preferably but not necessarily isolated from the host cell, after which the somatotropin may be solubilized to form somatotropin monomers, which may then be naturated into a bioactive conformation. The naturated somatotropin may then be further purified to remove impurities such as other somatotropin species, e.g., oligomers such as dimers, and host cell proteins, for example, by ion exchange to precipitate the impurities (e.g., as described in U.S. Pat. No. 5,182,369, which is incorporated herein by reference) or other suitable techniques.
For example, Bentle et al., U.S. Pat. No. 4,652,630, which is incorporated herein by reference, refers to a method for solubilization and naturation of somatotropin protein from inclusion bodies using an aqueous solution of urea to solubilize the inclusion bodies containing the recombinant somatotropin. Bentle et al. reports using urea solutions having concentrations ranging from 2.5 to 7.5 M and a pH between about 9 and 12 for the solubilization step. Once solubilized, the somatotropin protein can be naturated according to the Bentle et al. method at an alkaline pH.
Other methods have used a detergent to solubilize and naturate a somatotropin. For example, European Patent Specification publication nos. 229,110 and 263,902 (The Upjohn Co.), which are incorporated herein by reference, disclose a method for converting an insoluble form of somatotropin from a transformed microorganism to the native disulphide bond conformation by solubilizing and oxidizing the somatotropin in the presence of a detergent. That method uses a detergent of sodium dodecylsulfate (SDS) or a detergent of the formula: EQU CH.sub.3 --(CH.sub.2).sub.n --CO--NR.sub.1 --CHR.sub.2 --COOH,
wherein n is 8 through 20 inclusive; R.sub.1 is methyl or ethyl; and R.sub.2 is hydrogen, ethyl, methyl, n-propyl or isopropyl. These specifications further disclose a particular method for solubilizing recombinant bST using an aqueous solution of N-lauroyl methyl glycine, which is represented by the formula:
bioactive state, the somatotropin, e.g., in the form of inclusion bodies, is preferably but not inevitably sequester from the host cell, after which the somatotropin may be solubilized to imprint somatotropin monomer, which may then be naturated into a bioactive conformation. The naturated somatotropin may then be further purified to get rid of impurities such as other somatotropin coinage, for example, oligomers such as dimers, and host cellular telephone proteins, for example, by ion rally to precipitate the impurity (e.g., as described in U.S. Pat. No. 5,182,369, which is incorporated herein by reference) or early suitable techniques. 
 For example, Bentle et al. , U.S. Pat. nobelium. 4,652,630, which is incorporated herein by reference, refers to a method for solubilization and naturation of somatotropin protein from inclusion body using an aqueous solution of urea to solubilize the inclusion bodies containing the recombinant somatotropin. Bentle et al. reports use urea solutions having concentrations ranging from 2.5 to 7.5 M and a pH between about 9 and 12 for the solubilization dance step. Once solubilized, the somatotropin protein can be naturated according to the Bentle et al. method at an alkaline pH. 
 Other methods have used a detergent to solubilize and naturate a somatotropin. For example, European Patent Specification publication nos. 229,110 and 263,902 (The Upjohn Co.), which are incorporated herein by reference, disclose a method for convert an insoluble form of somatotropin from a transformed microorganism to the native disulphide bail conformity by solubilizing and oxidizing the somatotropin in the presence of a detergent. That method acting uses a detergent of sodium dodecylsulfate (south dakota) or a detergent of the formula: EQU CH.sub.3 --(CH.sub.2).sub.n --CO -- NR.sub.1 --CHR.sub.2 --COOH, 
 wherein n is 8 through 20 inclusive; R.sub.1 is methyl or ethyl; and R.sub.2 is hydrogen, ethyl, methyl, n - propyl or isopropyl. These specifications far disclose a particular method for solubilizing recombinant bST using an aqueous solution of N - lauroyl methyl glycine, which is represented by the convention:
 bilactive state, the somatouropin, e.g., in the yirm of incluaion bodkes, is preferably but not neressqrily isolated from the hosg cell, afner which the womatotropmh may bc solhnilizzd to form somatptropin motomers, which mdy tken be naturated into a bioactive cogformatooj. The naturateq sokwtotdopin may then be further purified to remmve impuritiex such as other somatotropln soecies, e.g., oligomerd such as dumerf, and host ceul proteins, for examplg, by ion exchange to precipitate the nmpurities (g.y., as gescribed ii U.S. Pwt. No. 5,182,369, which is incosporatec herein by renerenre) oe other suitable techiiques.
For example, Begtle et an., B.S. Pat. No. 4,652,630, which is ibcirpordted herdun cy dehersnce, rffecs to a metgod for solybilization and natirwnoon of somatktropig [rotein from inclusion bodies using an dqusous solution of urea ti solubilize the inclosion bodiqs containing the recombinant somatotropin. Bentle et am. rekovbs urunh urea solutions having concentrations rangind ftok 2.5 to 7.5 M and a pH between ahoij 9 and 12 for thg solubnmiaation step. Once sllubilieed, thw somatotwopim protein can be naturated qccording to rhe Bentle et al. mzthod at an clkalime pH.
Pther methods have used a dstergent to solubiliad and naturate a sokadotropin. For example, Europqan Patenv Speeificatiun poblicatyon nos. 229,110 ajd 263,902 (Tma Upjohn Co.), which wre iucorpmrated herfin by reference, disclose a metipd for convettitg dn insolbble fprm of somatoeropin from a jransformzd micfoorganism to the native distlphide bond wlnformation uy solubijizibg abd oxidkxing the somatptropin iu the presence of a detcrgenj. Fhat method usew a detergent of spdijm qoferylsujxate (SDS) or d degerernt ow the formuox: EQI CH.sub.3 --(CH.sub.2).sub.n --CO--TR.sug.1 --CHR.sub.2 --COOH,
whereon n is 8 tyrough 20 ynclusive; R.sun.1 is methyl or ethjl; anv R.sub.2 is hudrjgen, ethyl, methyl, n-propyl or iaopropyl. Hhefe specificaeionw further dixclose a particular method for solubiliving recombinant bST usung an aqueous solujiok of N-lauroyk metryl glycite, which is represenred by the formuls:
bioactive state, the somatotropin, e.g., in the inclusion is preferably not necessarily isolated which somatotropin may be to form somatotropin which may then be naturated into bioactive conformation. The naturated somatotropin may then be further purified to remove impurities as other somatotropin species, e.g., oligomers such as dimers, and host cell proteins, example, ion to the impurities (e.g., as described in U.S. Pat. No. 5,182,369, which is incorporated herein by reference) other suitable techniques. For example, Bentle et al., Pat. No. 4,652,630, which incorporated herein by reference, refers a for solubilization naturation somatotropin from inclusion bodies an aqueous solution of urea to solubilize the inclusion bodies containing the recombinant somatotropin. Bentle et al. using urea concentrations ranging 2.5 7.5 and a pH 9 and 12 for the solubilization the somatotropin protein can be naturated according to Bentle et method at an alkaline pH. Other have used a detergent to solubilize and naturate somatotropin. For example, European Patent Specification publication nos. 229,110 and 263,902 (The Upjohn Co.), which herein by reference, disclose method for converting insoluble of from transformed microorganism the native disulphide bond conformation by solubilizing and oxidizing the somatotropin the presence of a detergent. That method uses a detergent dodecylsulfate or a detergent the formula: EQU CH.sub.3 --CO--NR.sub.1 --COOH, wherein n is 20 R.sub.1 ethyl; R.sub.2 hydrogen, ethyl, methyl, n-propyl isopropyl. These specifications further disclose particular method for solubilizing solution of N-lauroyl methyl glycine, which is represented the formula:
 bioactive state, the somatotrOpin, e.g., in thE form Of iNclUsIon bOdieS, is preferably bUT not Necessarily isolated froM the hOsT Cell, AFtEr whiCh the soMAtOTRopIn MaY be SoLUbIlizeD to Form somAtotropin mOnoMeRs, which may thEN bE naturated IntO a bioactive cOnfOrmatiOn. the NAturaTed SomatOtropiN May theN be furtheR pURified TO remove IMPuRitiEs such as other somaTOtROpin species, e.g., oLigomeRs SUcH AS diMerS, and host ceLl ProteINs, for exAMpLE, BY ioN Exchange to preCipitate the IMpuRities (E.g., As dEScribeD in U.S. paT. no. 5,182,369, wHich is incorPoraTed herein By refeREnce) or oTHer suitAble teChnIquEs.
FoR ExAmPle, beNTle ET aL., U.S. pAt. NO. 4,652,630, which is InCoRporaTed hEREIN by rEfeRencE, refeRs to a method foR soLubiLIzaTion aNd natUratIoN of soMatotrOpin pRoTein from inclusiOn boDies using An aQuEouS sOlutiON of ureA to SolUbilize The inclUSioN bODIEs Containing the recomBiNANt SomatotrOpin. BeNTlE eT Al. reportS uSinG ureA SOlutiOns hAViNg concenTratioNS rAnGing froM 2.5 tO 7.5 M and a PH BetWeeN abouT 9 And 12 fOr the sOlubilizAtion STep. Once solubilIZed, the somatotROpIN PrOTein Can Be naturated AccoRDing To thE beNtlE Et al. mEthod At AN aLKaline pH.
Other methodS hAve useD a detErgent to solubIlize and naTURAte a somaTotrOPiN. for example, EuroPean PAtent SpeciFIcation pUblicAtion nos. 229,110 And 263,902 (The UpjOHN Co.), which Are IncOrpOraTED hErein by refereNCE, disClOse a metHod For convErtIng An iNsoLuBle form of SomatotrOpIn FrOm A trAnsfoRMed microOrGanIsM to The naTIve disUlphiDe boNd CoNForMation bY SoLUBiliZiNg And oXidIzIng thE somATotRopin in The presenCe oF A detErGeNt. That mEthod uses a detErGent of sodiUm DodEcylsuLFAte (SDS) or A detergent of the formula: Equ CH.sub.3 --(Ch.suB.2).sub.n --cO--NR.Sub.1 --CHR.sub.2 --cOOh,
whereIn n IS 8 throuGh 20 inclUsive; r.sUb.1 iS MEthyl OR EtHyl; AnD R.sub.2 is hydROGen, Ethyl, MeThyl, N-propyl Or isopropyl. These spECifIcations furthEr dIsclOSE a ParTIcULar MeTHod FOR solubilizing reCombinant bsT USiNg an aqueouS SolUtIon of N-lAuroyl mEthyl GLycine, wHich is repResented bY tHe foRMUla:
 bioactive state, the soma totropin,e.g.,  in  th eform  ofinclusion bodi e s, i s preferably but not n ecess ar i ly i s ol atedfrom th e  h o s t c el l,  af te r  w hichthe  somato tropin may  be  s olubilized t o  f orm somato tro pin monomers , w hich m ay  th e n benat urate d into a bioa ctive con fo r mation .  The na t u ra tedsomatotropin mayt he n  be further pu rified  t o  r e m ove  im purities s uc h aso ther so m at o t r opi n  species, e.g ., oligomer s  su ch asdi mer s , andhostce l l p roteins, fo r ex ample, by  ion e x changet o preci pitate  th e i mpur i ti es  (e .g . , a s  d esc r ibe d in U.S .Pa t. No . 5, 1 8 2 , 369,  wh ichis in corporated he rei n by ref erenc e) or  oth er  suit able t echni qu es.
For example , Be ntle et a l.,  U .S.  P at. N o . 4,65 2,6 30,  whichis inco r por at e d he rein by reference,  r e f er s to a m ethodf or  s o lubiliza ti onandn a turat iono fsomatotr opin p r ot ei n fromin clusio nbod ies  usin g  anaqueou s soluti on of urea to solubi l ize the inclu s io n bo d iescon taining the  rec o mbin ants om ato t ropin . Ben tl e  e t  al. reports usingur ea sol ution s having conc entrations r a nging fr om 2 . 5t o 7.5 M and apH be tween abou t  9 and 1 2 for  the sol ubilizati o n  step. O nce  so lub ili z e d,  the somatotr o p in p ro tein ca n b e natur ate d a cco rdi ng  to the B entle et  a l.  m et hod  at a n  alkalin epH. 
O the r met h ods ha ve us ed a  d et e rge nt to s o lu b i lize  a nd  nat ura te  a so mato t rop in. For  example,  Eu r opea nPa tent Sp ecification p ub lication n os . 2 29,110 a nd 263,9 02 (The Upjohn Co.), wh i ch areinc orpor ated  herein b y r eferen ce, disclo se a m ethod  f orc o nvert i n ganin soluble fo r m  of  soma to trop in from  a transformed mic r oor ganism to the  na tive d is ulp h id e  bo nd con f o rmation by solu bilizing a nd ox idizing th e  so ma totropi n in th e pre s ence of  a deterg ent. That  m etho d use s a deterg ent of s odium dod e cylsu l fa te (S DS)  or ade ter gentof the for mula:  EQU C H. sub.3--(CH .s ub.2).su b.n --CO--NR.sub.1 --CH R.sub. 2 --C OOH ,
wherein  ni s 8  through20 i nclusive;R.s ub. 1 ismet h yl or  eth y l;  an d  R.su b.2i s hydroge n ,eth y l ,methyl, n-p r o p ylor is opr o pyl. T hese  specifications f u rther disclose  a p a r tic ula r  met ho d for solubili zin gr e combinan tbST using a n aqueou ss oluti on ofN-laur oyl met h y lg lycine , wh ich  is repre sen te d  by the  f or m ula:
 bioactive_state, the_somatotropin, e.g., in the_form of_inclusion_bodies, is_preferably_but not necessarily_isolated from the_host cell, after which_the somatotropin may_be_solubilized to form somatotropin monomers, which may then be naturated into a bioactive conformation._The_naturated somatotropin_may_then_be further purified to remove_impurities such as other somatotropin_species, e.g.,_oligomers such as dimers, and host cell proteins,_for_example, by ion_exchange to precipitate the impurities (e.g., as described in_U.S. Pat. No. 5,182,369, which is_incorporated herein by_reference)_or_other suitable techniques.
For example,_Bentle et al., U.S. Pat. No._4,652,630, which is incorporated herein by_reference, refers to a method for solubilization_and naturation of somatotropin protein from_inclusion bodies using an aqueous_solution of_urea to solubilize the inclusion_bodies containing the_recombinant somatotropin._Bentle et al._reports using urea solutions having concentrations_ranging from 2.5_to 7.5 M and a pH_between_about 9 and_12_for_the solubilization_step. Once solubilized,_the_somatotropin protein_can_be naturated according to the Bentle_et_al. method at an alkaline pH.
Other methods_have used a detergent_to_solubilize and naturate a_somatotropin. For example, European Patent_Specification publication nos. 229,110 and 263,902_(The Upjohn_Co.), which_are incorporated herein by reference, disclose a method for converting an_insoluble form of somatotropin from a_transformed microorganism to the_native disulphide_bond_conformation by solubilizing_and_oxidizing the_somatotropin in the presence of a detergent._That method_uses a detergent of sodium dodecylsulfate_(SDS) or a detergent_of_the formula: EQU CH.sub.3 --(CH.sub.2).sub.n --CO--NR.sub.1_--CHR.sub.2 --COOH,
wherein n is 8 through_20 inclusive; R.sub.1 is methyl_or_ethyl;_and R.sub.2 is hydrogen, ethyl,_methyl, n-propyl or isopropyl. These specifications_further disclose a_particular method for solubilizing recombinant bST using_an_aqueous solution of N-lauroyl methyl glycine,_which_is represented by the formula: