Antibody Engineering ni iki?

Hifashishijwe ubuhanga bwa antibody, byashobokaga guhindura ingano ya molekile, pharmacokinetics, immunogenicite, guhuza isano, umwihariko hamwe nibikorwa bya antibodies. Nyuma yo guhuza antibodi, guhuza antibodi byihariye bituma bigira agaciro gakomeye mugupima no kuvura. Binyuze muri antibody yubuhanga, barashobora guhaza ibikenerwa nibiyobyabwenge no kwisuzumisha hakiri kare.

Intego yubuhanga bwa antibody nugushushanya no gutanga umusaruro wihariye, uhamye antibodi karemano idashobora kugeraho, igashyiraho urufatiro rwo gukora antibodiyite zivura.

Alpha Lifetech, hamwe nuburambe bunini bwumushinga mu buhanga bwa antibody, irashobora gutanga serivisi yihariye ya antibody ya monoclonal na polyclonal antibody yubwoko butandukanye, hamwe na fage yerekana ububiko bwibitabo bwububiko bwa serivisi hamwe no gusuzuma. Alpha Lifetech irashobora guha abakiriya antibodiyite nziza ya biosimilar hamwe nibicuruzwa bya poroteyine bya recombinant, hamwe na serivisi zijyanye nabyo, kugirango bibyare antibodi nziza, zihariye, kandi zihamye. Dukoresheje antibody yuzuye, porogaramu za poroteyine hamwe na sisitemu yo kwerekana ibyiciro, dutanga serivisi zikubiyemo epfo na ruguru y’umusemburo wa antibody, harimo serivisi za tekiniki nko kuba antibody ya kimuntu, kweza antibody, gukurikirana antibody, no kwemeza antibody.

Icyiciro cya mbere cyubwubatsi bwa antibody kijyanye nubuhanga bubiri:

--Ikoranabuhanga rya ADN

- Ikoranabuhanga rya Hybridoma

Iterambere ryihuse ryubwubatsi bwa antibody rifitanye isano na tekinoroji eshatu zingenzi:

--Gene clone tekinoroji hamwe na polymerase urunigi

- Imvugo ya poroteyine: Poroteyine za Recombinant zikorwa na sisitemu yo kwerekana nk'umusemburo, virusi zimeze nk'inkoni, n'ibimera.

--Mudasobwa yafashijwe igishushanyo mbonera

Igisekuru cya mbere cya antibodiyite zahinduwe abantu kugirango bakore antibodiyite za chimeric, aho akarere kahinduye imbeba ya monoclonal antibodies zahujwe nakarere gahoraho ka molekile zabantu. Agace ka antigen gahuza (CDR) ya antibody ya kabiri yimbeba ya monoclonal antibody yatewe muri IgG yabantu. Usibye akarere ka CDR, izindi antibodiyose zose ni antibodi zabantu, kandi hashyizweho ingufu kugirango birinde gutera antibody anti-antibody yumuntu (HAMA) mugihe ukoresheje antibodi ya clone yimbeba mukuvura abantu.

 

Kubaka isomero ryerekana fage, intambwe yambere nukubona gen zigizwe na antibodies, zishobora gutandukanywa na selile B yinyamaswa zakingiwe (kubaka isomero ry’ubudahangarwa), zivanwa mu buryo butaziguye n’inyamaswa zidakingiwe (kubaka isomero karemano), cyangwa se ziteranirizwa muri vitro hamwe n’ibice bya gene (kubaka isomero ryububiko). Noneho, ingirabuzimafatizo zongerwaho na PCR, zinjizwa muri plasmide, kandi zigaragarira muri sisitemu ibereye (imvugo y’imisemburo (ubusanzwe Pichia pastoris), imvugo ya prokaryyotike (ubusanzwe E. coli), imvugo y’inyamabere y’inyamabere, imvugo y’ibimera, n’imvugo y’udukoko twanduye virusi zifite inkoni). Igikunze kugaragara cyane ni E. coli imvugo ya sisitemu, ihuza uburyo bwihariye bwa kodegisi ya antibody ikurikirana kuri fage kandi igahuza imwe muri poroteyine zo mu bwoko bwa page (pIII cyangwa pVIII). Gene fusion ya, Kandi yerekanwe hejuru ya bagiteri. Intandaro yikoranabuhanga ni ukubaka fage yerekana isomero, rifite inyungu kurenza amasomero karemano kuko rishobora kugira umwihariko. Nyuma, antibodies zifite umwihariko wa antigen zirasuzumwa binyuze muburyo bwo gutoranya ibinyabuzima, antigene zigamije gukosorwa, ibyiciro bitagira umupaka byogejwe inshuro nyinshi, kandi ibyiciro bihambirijwe byogejwe kugirango birusheho gukungahaza. Nyuma yincuro eshatu cyangwa nyinshi zisubiramo, umwihariko wo hejuru hamwe na antibodi nyinshi zifitanye isano.

Igishushanyo 3: Kubaka Isomero rya Antibody no Kugaragaza

Tekinoroji ya ADN ya recombinant irashobora gukoreshwa mugukora ibice bya antibody. Antibodiyite za Fab zirashobora kubanza gusa hydrolyz na protease ya gastricike kugirango itange (Fab ') ibice 2, hanyuma bigogorwa na papain kugirango bibyare ibice bya Fab. Igice cya Fv kigizwe na VH na VL, zifite umutekano muke kubera kubura inkwano ya disulfide. Kubwibyo, VH na VL bihujwe hamwe binyuze muri peptide ngufi ya aside amine 15-20 kugirango ikore antibody imwe yumunyururu (scFv) ifite antibody ifite uburemere bwa molekile hafi 25KDa.

Ubushakashatsi ku miterere ya antibody muri Camelidae (Ingamiya, LIama, na Alpaca) bwasobanuye ko antibodiyite zifite iminyururu iremereye gusa kandi nta munyururu woroheje, bityo bakaba bita antibodi ziremereye (hcAb). Imiterere ihindagurika ya antibodi iremereye yitwa antibodi imwe ya domeni cyangwa nanobodies cyangwa VHH, hamwe na 12-15 kDa. Nka ba monomers, nta sano ya disulfide bafite kandi irahagaze neza, hamwe cyane na antigene.

Igishushanyo 5: Antibody Yumunyururu Ukomeye na VHH / Nanobody

Imikorere yubusa ikoresha imvugo ya ADN karemano cyangwa sintetike kugirango igere muri synthesis ya vitro protein, mubisanzwe ikoresha sisitemu ya E. coli. Itanga poroteyine vuba kandi ikirinda umutwaro wa metabolike na cytotoxic kuri selile iyo utanga proteine ​​nyinshi za recombinant muri vivo. Irashobora kandi gukora poroteyine zigoye guhuza, nk'izigoye guhindura nyuma yo guhindurwa cyangwa guhuza poroteyine za membrane.