
taandam affinity shudheekaranam

protein-protein edapedalukal padikkunnathinulla rogaprathirodhasheshi adisthaanamaakkiyulla shudheekarana saankethikathayaanu tandom affinity purification (tapp). oru koshatthil ninnu thaalpparyamulla protein maathram verthirichedukkuka ennathaanu lakshyam, athu edapedunna mattethenkilum proteinukalumaayi sankeernamaayi. thaalpparyamulla proteinumaayi bandhippikkunna rund tharam agaros muthukal tapp upayogikkunnu, avaa sell laisettil ninnu centrifugation vazhi verthirikkaan kazhiyum, ulppettirikkunna complexukale shalyappeduthukayo apakeerthippeduthukayo malinappeduthukayo cheyyaruthu. thaalpparyamulla proteine muthukalumaayi bandhippikkaan praapthamaakkunnathin, athine tapp tagg enna roopakalppana cheytha kashanam upayogichu tagg cheiyunnu. 
yadhaartha tapp reethiyil padanatthilulla proteente sii-terminessilekku tapp tagg samyojippikkunnath ulppedunnu. tapp taggle moonnu gadakangal adangiyirikkunnu oru calamoduline binding pepttaid (cbp), tevu protees kleevej saitt, rund protein eiii domainukal, avaa iggumaayi karsanamaayi bandhippikkunnu (oru tapp tagg oru tharam epitoppu tagg akkunnu).
tapp thathwam aadyamaayi prasidheekarichathumuthal mattu niravadhi tagg/beed/eluent kombineshanukal nirddeshikkappettittund.
vyathyastha taagukal.
oru enn-terminal pathipum labhyamayathinaal eee tagg sii-terminal tapp tagg ennum ariyappedunnu. ennirunnaalum, vivarikkenda reethi oru sii-terminal taaginte upayogamaanennu anumaanikkunnu, ennirunnaalum eee reethikk pinnile thathwam ippozhum athepadi thanneyaanu.
charithram.
1990 kalude avasaanathil europian molecular biology laboratariyil pravartthikkunna oru gaveshana samghamaanu tapp tagging kandupidichathu.1999, puig thudangiyavar., 2001) proteom paryaveshanathinulla oru puthiya upakaranamaayi nirddeshichu. niravadhi protein complexukal (regout thudangiyava) savisheshamaakkaan team ithu upayogichu.1999, kaspari thudangiyavar. 1999, boovartt thudangiyava., 2000, puig thudangiyavar.2001). eee saankethikavidyayude aadyathe valia thothilulla prayogam 2002-layirunnu, athil proteomics kambaniyaaya selsominte shaasthranjarumaayi sahakarichu gaveshana samgham oro proteinilekkum tapp tagg chittayayi tagg cheythukondu oru yeest sellile 230-ladhikam multy-protein complexukalude prathipravartthanatthinte vishwal mapp vikasippichu. sasyangalil tapp tagg saankethikavidya upayogichathinte aadya vijayakaramaya report 2004-laanu (rohila mattullavarum) vannath.2004)
prakriya.
aathidheya koshangalil fusion protein avatharippikkan chila reethikalundu. aathidheyan yeest aanenkil, plaasmidukalude upayogamayirikkaam oru reethi, athu oduvil aathidheyarkkullile fusion proteine vivarthanam cheyyum. eathu reethiyaanu upayogikkunnathenkilum, fusion proteente aavishkaaram athinte swaabhaavika nilavaarathodu kazhiyunnathra aduthu nilanirthunnathaanu nallath. fusion protein hosttinullil vivarthanam cheythukazhinjaal, athu mattu proteinukalumaayi samvadikkum, tapp tagg baadhikkaatha reethiyil. 
pinneedu, tagg cheytha protein (athinte binding pankaalikalkoppam) oru affinity selection prakriya upayogichu veendedukkunnu.
tapp taaginte ettavum puram attathu bandhippikkunna immunoglobulin jii kondu pothinjathaanu aadyathe tharam beed cherkkunnath. thaalpparyamulla proteinukalulla muthukal centrifugation vazhi laisettil ninnu verthirikkunnu. proteinukal pinneedu oru ensime (tevu protees) kondu muthukalil ninnu purathuvidunnu, ithu madhyathilulla tevu pilarpp saittil tagg thakarkkunnu.
eee aadya shudheekarana ghattathinu sesham, puratthirakkiya proteinukalil randaamathe tharam beed (calamoduline pooshiya) cherkkunnu, ithu proteinukalil ippozhum tapp taaginte sheshikkunna bhagavumaayi thirichupokunnu. manikal veendum centrifugation vazhi verthirikkappedunnu, ithu malineekaranavasthukkal tevu protease kooduthal neekkam cheiyunnu. oduvil, muthukal egta vazhi purathuvidunnu, thaalpparyamulla protein, athinte bandhita protein pankaalikal, tapp taaginte bakkiyulla cbp kashanam enniva maathram adangiyirikkunna native eluyettu avasheshikkunnu.
proteente binding pankaalikale thirichariyaan jell electrophoresis maas spectrometry upayogichu native eluyettu vishakalanam cheyyaan kazhiyum.
nettangal.
sankeernamaaya gadanayekkurichu munkootty ariyaathe vivoyil protein pankaalikalude alavu yadhaarthathil nirnnayikkaan kazhiyum ennathaanu eee reethiyude oru nettam. ithu nadappilakkaanum eluppamaanu, palappozhum uyarnna vilavu nalkunnu. protein protein edapedal padikkunnathinulla thadassangalilonnu taargett protein malineekaranamaanu, prathyekichu namukk athinekkurichu munkootty arivillaathappol. tapp taargett protein shudheekarikkunnathinula phalapradavum valare nirddishtavumaaya maarggam vaagdaanam cheiyunnu. thudarchayaayi 2 affinity shudheekaranangalkku sesham, malineekaranavasthukkal eluvettil nilanirthaanulla saadhyatha ganyamaayi kurayunnu.
poraaymakal.
ennirunnaalum, oru proteinil cherttha oru tagg puthiya proteine athinte paraspara pankaalikalumaayi bandhippikkunnath avyakthamakkaanum saadhyathayund. koodaathe, tagg protein expression levelukaleyum baadhichekkaam. maruvashathu, tagg affinity muthukalkku vendathra thurannukaattappedaathirim, athinaal phalangal valayunnu.
tevu proteesinte uyarnna prathyekatha kanakkiledukkumbol ithu pathivayi sambhavikkaan saadhyathayillengilum tevu protees proteinukalude pilarppinu saadhyathayund.
anuyojyatha.
eee reethiyil kuranjathu 2 raundu kazhukal ulppedunnathinaal, yeest tuu-hibrid reethi allengil foto-reactive amino assid analogukalumaayi cross linking cheiyunna "inn vivo" ennathil ninnu vyathyasthamayi, shanikamaaya protein edapedalukal parisodhikkaan ithu anuyojyamalla. ennirunnaalum, sthirathayulla protein edapedalukal parishodhikkunnathinulla oru nalla reethiyaanithu, koodaathe protein complexes ethra thavana shudheekarikkappedunnu ennu niyanthrikkunnathiloode vividha thalatthilulla anweshanam anuvadikkunnu.
aaplikkeshanukal.
multiprotein complexukalude swabhaavasavisheshathakal vyakthamaakkikkond yeesttinte proteomics chittayayi vishakalanam cheiyunnathinaayi 2002-ll maas spectrometry upayogichu valia thothilulla sameepanathilaan tapp tagg aadyamaayi upayogichathu. 257 complexukal poornnamaayum puthiyavayaya 491 complexukal padanatthil kandethi. bakkiyullava mattu gaveshanangalil ninnu parichithamaayirunnu, ennaal ippol avaykku mikkavarum ellah puthiya gadakangalum undennu kandethi. sankeernamaaya oru shringalayile ellah protein gadakangaleyum sambandhicha oru bhoopadam avar thayyarakki.
mattu pala proteomic vishakalanangalilum tapp taaginte upayogam ulppedunnu. embo (dii. seembowski, 2004) nadatthiya oru gaveshanathil nuclear pree-marnah nilanirthunnathinum spliesingin aavashyamaaya oru puthiya samuchayam kandethi. mattu 3 subunitukal (snu 17pii, budd 13pii, pml 1pii) ulkkollunna oru puthiya trimeric complexes avar shudheekarikkukayum eee subunitukal pravarthanakshamathaykku anivaaryamallennum ennaal pree-marnayude kaaryakshamamaya spliesingin (intronukal neekkam cheiyunnathinu) aavasyamaanennum kandethukayum cheythu. 2006-ll, "fleesher mattullavarum."eucariotic ribosomal komplexukalumaayi bandhapetta proteinukal vyavasthaapithamaayi thiricharinju. yeest ribosomal complexukal thirichariyaan avar bahumukha maas spectrometry proteomic screenukal upayogikkukayum thudarnnu eee proteinukalellaam pravartthanaparamaayi bandhippikkunnathin tapp tagging upayogikkukayum cheythu.
mattu epitoppu-tagg kombineshanukal.
multiprotein complexukalude tandom-affinity shudheekaranathinte thathwam sii. bii. pii, protein ennivayude samyojanathil parimithappedunnilla-regout thudangiyavar yadhaartha srishtiyil upayogicha taagukal. (1999). udaaharanathinu, sasthanikalude koshangalil ninnu niravadhi protein complexukal shudheekarikkaan 2000 muthal nakadaani group flag-haah-taagukalude samyojanam upayogikkunnu. tapp thathwam prasidheekarichathinushesha mattu niravadhi tagg kombineshanukal nirddeshikkappettittund.

