Kuyini Ubunjiniyela Bokulwa Nomzimba?

Ngosizo lobunjiniyela be-antibody, kuye kwenzeka ukushintsha usayizi wama-molecule, i-pharmacokinetics, i-immunogenicity, i-binding affinity, i-specificity kanye nomsebenzi we-effector wama-antibodies. Ngemva kokwenza ama-antibodies, ukubopha okuthile kwama-antibodies kuwenza abe usizo kakhulu ekuxilongweni nasekwelashweni kwezokwelapha. Ngobunjiniyela be-antibody, angahlangabezana nezidingo zokuthuthukiswa kwemithi kanye nokuxilongwa kusenesikhathi.

Inhloso yobunjiniyela bama-antibody ukuklama nokukhiqiza imisebenzi eqondile kakhulu nezinzile ama-antibody emvelo angenakuyifeza, okubeka isisekelo sokukhiqizwa kwama-antibody okwelapha.

I-Alpha Lifetech, enokuhlangenwe nakho kwayo okukhulu kwephrojekthi yobunjiniyela be-antibody, ingahlinzeka ngezinsizakalo ze-antibody ze-monoclonal neze-polyclonal ezenzelwe wena zezinhlobo eziningi, kanye nezinsizakalo zokwakha kanye nokuhlola umtapo wolwazi we-antibody ye-phage display. I-Alpha Lifetech ingahlinzeka amakhasimende ngama-antibodies e-biosimilar asezingeni eliphezulu kanye nemikhiqizo yamaprotheni ahlanganisiwe, kanye nezinsizakalo ezifanayo, ukukhiqiza ama-antibodies asebenzayo, aqondile kakhulu, futhi azinzile. Ngokusebenzisa ama-antibody aphelele, amapulatifomu amaprotheni kanye nezinhlelo zokubonisa i-phage, sinikeza izinsizakalo ezimboza ukukhiqizwa kwama-antibody okuphezulu nangaphansi, kufaka phakathi izinsizakalo zobuchwepheshe ezifana nokwenza ama-antibody abe ngabantu, ukuhlanzwa kwama-antibody, ukulandelana kwama-antibody, kanye nokuqinisekiswa kwama-antibody.

Isigaba sokuqala sobunjiniyela bama-antibody sihlobene nobuchwepheshe obubili:

--Ubuchwepheshe be-DNA obuhlanganisiwe

--Ubuchwepheshe be-Hybridoma

Ukuthuthukiswa okusheshayo kobunjiniyela bama-antibody kuhlobene nobuchwepheshe obuthathu obubalulekile:

--Ubuchwepheshe bokuklonyela izakhi zofuzo kanye nokusabela kweketanga le-polymerase

--Ukubonakaliswa kwamaprotheni: Amaprotheni aphinde ahlanganiswa akhiqizwa izinhlelo zokubonakaliswa njengemvubelo, amagciwane anomumo wenduku, kanye nezitshalo

--Umklamo wesakhiwo osizwa yikhompyutha

Isizukulwane sokuqala sama-antibodies senziwe saba ngabantu ukuze kukhiqizwe ama-antibodies e-chimeric, lapho indawo eguquguqukayo yama-antibodies e-monoclonal yegundane yayixhunyaniswe nendawo engaguquki yama-molecule e-IgG yomuntu. Isifunda sokubopha ama-antigen (i-CDR) se-antibody ye-monoclonal yegundane yesizukulwane sesibili satshalwa ku-IgG yomuntu. Ngaphandle kwesifunda se-CDR, wonke amanye ama-antibodies cishe angama-antibodies abantu, futhi kwenziwa imizamo yokugwema ukuthonya izimpendulo zama-antibody e-anti yegundane lomuntu (i-HAMA) lapho kusetshenziswa ama-antibodies e-clone yegundane ekwelapheni abantu.

 

Ukuze wakhe umtapo wolwazi we-phage, isinyathelo sokuqala ukuthola izakhi zofuzo ezifaka ama-antibodies, ezingahlukaniswa kumaseli e-B ezilwane ezigonyiwe (ukwakhiwa komtapo wolwazi we-immune), ezikhishwe ngqo ezilwaneni ezingagonyiwe (ukwakhiwa komtapo wolwazi wemvelo), noma zihlanganiswe nge-vitro ngezingcezu zezakhi zofuzo ze-antibody (ukwakhiwa komtapo wolwazi wemvelo). Ngemuva kwalokho, izakhi zofuzo ziyandiswa yi-PCR, zifakwe kuma-plasmid, futhi ziboniswe ezinhlelweni ezifanele zokusingatha (ukubonakaliswa kwemvubelo (ngokuvamile i-Pichia pastoris), ukubonakaliswa kwe-prokaryotic (ngokuvamile i-E. coli), ukubonakaliswa kwamaseli ezilwane ezincelisayo, ukubonakaliswa kwamaseli ezitshalo, kanye nokubonakaliswa kwamaseli ezinambuzane atheleleke ngamagciwane anomumo wenduku). Okuvame kakhulu uhlelo lokubonakaliswa kwe-E. coli, oluhlanganisa ukulandelana okuthile kwe-antibody ekhodi ku-phage futhi lufaka ikhodi kwenye yamaprotheni egobolondo le-phage (pIII noma i-pVIII). Ukuhlanganiswa kwezakhi zofuzo, Futhi kuboniswa ebusweni be-bacteriophages. Ingqikithi yalobu buchwepheshe ukwakha umtapo wolwazi we-phage, onenzuzo ngaphezu kwemitapo yolwazi yemvelo ngoba ungaba nokubopha okuthile. Ngemva kwalokho, ama-antibodies ane-antigen specificity ahlolwa ngenqubo yokukhetha izinto eziphilayo, ama-antigen aqondiwe ayalungiswa, ama-phage angaboshiwe ahlanzwa ngokuphindaphindiwe, futhi ama-phage aboshiwe ayahlanzwa ukuze kuthuthukiswe okwengeziwe. Ngemva kokuphindaphinda kathathu noma ngaphezulu, ama-antibodies ane-specificity ephezulu kanye ne-high affinity ayahlukaniswa.

Umfanekiso 3: Ukwakhiwa Nokuhlolwa Komtapo Wolwazi We-Antibody

Ubuchwepheshe be-Recombinant DNA bungasetshenziswa ukukhiqiza izingcezu zama-antibody. Ama-antibody e-Fab ekuqaleni angafakwa kuphela yi-gastric protease ukuze kukhiqizwe izingcezu ezi-2 (Fab '), ezibe sezigaywa yi-papain ukuze kukhiqizwe izingcezu ze-Fab ngazinye. Isingcezu se-Fv sakhiwe yi-VH ne-VL, ezingakwazi ukuzinza kahle ngenxa yokungabikho kwezibopho ze-disulfide. Ngakho-ke, i-VH ne-VL zixhunyaniswa ndawonye nge-peptide emfushane yama-amino acid angu-15-20 ukuze kwakhiwe i-antibody ye-single chain variable fragment (scFv) enesisindo sama-molecule esingaba ngu-25KDa.

Ucwaningo lwesakhiwo se-antibody ku-Camelidae (Camel, LIama, kanye ne-Alpaca) lucacise ukuthi ama-antibodies anezintambo ezisindayo kuphela futhi awanazo izintambo ezilula, yingakho ebizwa ngokuthi ama-antibody ezintambo ezisindayo (hcAb). Isizinda esiguquguqukayo sama-antibody ezintambo ezisindayo sibizwa ngokuthi ama-antibody e-single domain noma ama-nanobodies noma i-VHH, anosayizi ongu-12-15 kDa. Njengama-monomers, awanazo izibopho ze-disulfide futhi azinzile kakhulu, anobuhlobo obuphezulu kakhulu nama-antigen.

Umfanekiso 5: I-Antibody Ene-Heavy Chain kanye ne-VHH/ Nanobody

Ukukhululeka kweseli kusebenzisa ukuvezwa kwe-DNA yemvelo noma yokwenziwa ukuze kufezwe ukwakheka kwamaprotheni e-in vitro, ngokuvamile kusetshenziswa uhlelo lokuvezwa kwe-E. coli. Ikhiqiza amaprotheni ngokushesha futhi igwema umthwalo we-metabolic kanye ne-cytotoxic kumaseli lapho ikhiqiza inani elikhulu lamaprotheni aphinde ahlanganiswe emzimbeni. Ingakhiqiza futhi amaprotheni okunzima ukuwahlanganisa, njengalawo okunzima ukuwaguqula ngemva kokuguqulwa noma ukuhlanganiswa kwamaprotheni e-membrane.