Kedu ihe bụ Antibody Engineering?

Site n'enyemaka nke injinia antibody, enwere ike gbanwee nha molekul, pharmacokinetics, immunogenicity, njikọ njikọ, nkọwapụta na arụ ọrụ mmetụta nke ọgwụ nje. Mgbe emechara ọgwụ mgbochi ọrịa, njide kpọmkwem nke ọgwụ nje na-eme ka ha baa uru nke ukwuu na nyocha na ọgwụgwọ ụlọ ọgwụ. Site na injinia antibody, ha nwere ike gboo mkpa ọgwụ na nchọpụta mmalite mmalite.

Ebumnuche nke injinia mgbochi mmadụ bụ imepụta na imepụta ọrụ kwụsiri ike nke mgbochi anụ ahụ enweghị ike ịrụzu, na-atọ ntọala maka mmepụta ọgwụ mgbochi ọrịa.

Alpha Lifetech, yana ahụmịhe ọrụ ya buru ibu na injinia mgbochi mmadụ, nwere ike ịnye ọrụ mgbochi monoclonal ahaziri iche na polyclonal maka ọtụtụ ụdị yana phage ngosi ihe owuwu ụlọ ọba akwụkwọ yana ọrụ nyocha. Alpha Lifetech nwere ike inye ndị ahịa ọgwụ mgbochi biosimilar dị mma yana ngwaahịa protein recombinant, yana ọrụ kwekọrọ, iji mepụta ọgwụ mgbochi dị mma, akọwapụtara nke ọma na kwụsiri ike. Site n'iji ọgwụ mgbochi zuru oke, nyiwe protein na sistemu ngosipụta phage, anyị na-enye ọrụ na-ekpuchi elu na mgbada nke mmepụta ihe mgbochi, gụnyere ọrụ nka dị ka mgbochi mmadụ, ịdị ọcha mgbochi, usoro mgbochi mmadụ, yana nkwenye mgbochi.

Usoro ọsụ ụzọ nke injinia antibody metụtara teknụzụ abụọ:

--Teknụzụ DNA recombinant

-- teknụzụ hybridoma

Mmepe ngwa ngwa nke injinia antibody metụtara teknụzụ atọ dị mkpa:

--Gene cloning technology na polymerase chain mmeghachi omume

--okwu Protein: A na-emepụta protein ndị na-emegharị emegharị site na usoro okwu dị ka yist, nje ndị yiri mkpanaka, na osisi.

--Nhazi nhazi nke kọmpụta nyere aka

Ọgbọ nke mbụ nke ọgwụ mgbochi bụ mmadụ maka imepụta ọgwụ mgbochi chimeric, ebe mpaghara mgbanwe nke nje monoclonal òké jikọtara ya na mpaghara ụmụ irighiri ihe IgG mgbe niile. A tụgharịrị mpaghara antigen binding (CDR) nke ọgbọ nke abụọ na-egbochi monoclonal òké n'ime mmadụ IgG. Ewezuga mpaghara CDR, ọgwụ mgbochi ndị ọzọ niile fọrọ nke nta ka ọ bụrụ ọgwụ mgbochi mmadụ, a gbakwara mbọ iji zere ịkpata mmeghachi omume mgbochi mgbochi ụmụ mmadụ (HAMA) mgbe ị na-eji ọgwụ mgbochi òké maka ọgwụgwọ mmadụ.

 

Iji wuo ọbá akwụkwọ ihe ngosi nke phage, nzọụkwụ mbụ bụ inweta mkpụrụ ndụ ihe nketa na-edobe ọgwụ mgbochi ọrịa, bụ́ nke a pụrụ ikewapụ n’ime mkpụrụ ndụ B nke anụmanụ ndị a na-alụso ọrịa ọgụ (iwu ọbá akwụkwọ na-alụso ọrịa ọgụ), nke a na-amịpụta kpọmkwem site n’anụmanụ ndị na-abụghị ọgwụ mgbochi (iwu ọbá akwụkwọ nkịtị), ma ọ bụ ọbụna gbakọta na vitro nwere iberibe mkpụrụ ndụ ihe mgbochi (ihe owuwu ọbá akwụkwọ synthetic). Mgbe ahụ, a na-amụba mkpụrụ ndụ ihe nketa site na PCR, tinye ya n'ime plasmids, ma gosipụta ya na usoro ndị ọbịa kwesịrị ekwesị (okwu yist (na-abụkarị Pichia pastoris), okwu prokaryotic (na-emekarị E. coli), mkpụrụ ndụ anụ ahụ mammalian, okwu mkpụrụ ndụ osisi, na mkpụrụ ndụ ụmụ ahụhụ na-ebute nje ndị na-eme mkpara). Nke a na-ahụkarị bụ sistemu okwu E. coli, nke na-ejikọta usoro nzuzo nzuzo nzuzo na phage ma tinye otu n'ime protein phage shell (pIII ma ọ bụ pVIII). The gene fusion nke, Na-egosipụta n'elu bacteriophages. Isi ihe dị na teknụzụ a bụ ịrụ ụlọ akwụkwọ ngosi phage, nke nwere uru karịa ọba akwụkwọ okike n'ihi na ọ nwere ike ịnwe njikọ akọwapụtara. Mgbe nke ahụ gasịrị, a na-enyocha ọgwụ mgbochi ọrịa nwere ụdị antigen site na usoro nhọrọ nke ndu, na-edozi antigens ezubere iche, na-asachapụ phages ndị a na-ekekọtaghị ugboro ugboro, na-asachapụkwa phages ndị a kekọtara maka ịba ụba. Mgbe ugboro atọ ma ọ bụ karịa ugboro ugboro ugboro ugboro, a na-ekewapụ ihe mgbochi dị elu na ihe mgbochi dị elu.

Nkeji 3: Mwube na nyocha ọbá akwụkwọ Antibody

Enwere ike iji teknụzụ DNA recombinant mepụta iberibe mgbochi mmadụ. Ọgwụ mgbochi ọrịa nwere ike ibido naanị hydrolyzed site na gastric protease iji mepụta (Fab ') 2 iberibe, nke papain na-agbari wee mepụta iberibe Fab n'otu n'otu. Iberibe Fv nwere VH na VL, nke nwere nkwụsi ike na-adịghị mma n'ihi enweghị nkekọ disulfide. Ya mere, VH na VL na-ejikọta ọnụ site na peptide dị mkpirikpi nke 15-20 amino acids iji mepụta otu ihe mgbochi mgbanwe (scFv) nke nwere ihe dị arọ nke ihe dịka 25KDa.

Ọmụmụ ihe gbasara usoro mgbochi mmadụ na Camelidae (Camel, LIama, na Alpaca) akọwapụtala na ọgwụ nje nwere naanị ụdọ dị arọ na enweghị ụdọ ọkụ, ya mere a na-akpọ ha ọgwụ mgbochi yinye dị arọ (hcAb). A na-akpọ ngalaba na-agbanwe agbanwe nke ọgwụ mgbochi yinye dị arọ otu ngalaba ọgwụ mgbochi ma ọ bụ nanobodies ma ọ bụ VHH, nwere nha 12-15 kDa. Dị ka monomers, ha enweghị njikọ nke disulfide ma kwụsie ike, na-enwe mmekọrịta dị elu maka antigens.

Fig 5: Nnukwu Chain Antibody na VHH/ Nanobody

Okwu nnwere onwe cell na-eji okwu nke DNA eke ma ọ bụ nke sịntetịt iji nweta njikọ protein vitro, na-ejikarị usoro okwu E. coli. Ọ na-emepụta protein ngwa ngwa ma na-ezere ibu arọ nke metabolic na cytotoxic na sel mgbe ọ na-emepụta nnukwu proteins recombinant na vivo. Ọ nwekwara ike ịmepụta protein ndị siri ike ịmepụta, dị ka ndị na-esi ike ịgbanwe mgbe nsụgharị ma ọ bụ synthesize protein protein.