Ngo-1960, kwaphakanyiswa umqondo wama-antibodies e-bispecific. Ama-antibodies e-bispecific, aziwa nangokuthi ama-antibodies e-bispecific monoclonal, angama-antibodies akhiwe ngokwenziwa kusetshenziswa ubuchwepheshe bobunjiniyela bezakhi zofuzo. Njenge-antibody yokwenziwa eyenziwe ngobunjiniyela, ama-antibodies e-bispecific ngokuvamile ayingxenye ye-IgG futhi aqukethe ingxenye yokubopha i-antigen eqondise i-subunit ye-CD3. Ama-antibodies e-bispecific anezindawo ezimbili ezithile zokubopha i-antigen ezingabopha futhi zibone ama-antigen amabili ahlukene ngasikhathi sinye, noma ama-epitopes amabili ahlukene e-antigen. Uma kuqhathaniswa nama-antibodies e-monoclonal, ama-antibodies e-bispecific anesinye isayithi sokubopha i-antigen esithile, ngaleyo ndlela abe nokucaciswa okuqinile nekhono lokuqondisa, okungaqondisa ngokunembile amaseli esimila futhi kuncishiswe ubuthi obungaqondiwe. Ama-antibodies e-bispecific angenza imisebenzi eminingi yebhayoloji ngasikhathi sinye, njengokuqasha amaseli omzimba, ukuvimba izindlela zokubonisa izimpawu, nokubulala amaseli esimila ngokuqondile. Ama-antibodies okuqala e-bispecific alungiswa kakhulu ngokuhlanganiswa kwamakhemikhali noma ukuhlanganiswa kwamaseli, kodwa le ndlela kungenzeka ukuthi yathuthuka kancane ngenxa yokuhlanganiswa okungahleliwe kanye nobunzima bokuhlukanisa inhlanganisela yokuqondisa. Ngokuthuthuka okuqhubekayo kobuchwepheshe bobunjiniyela bezakhi zofuzo, kuye kwasungulwa amapulatifomu amaningi amasha obuchwepheshe, njenge-knots in holes (KIH), i-CrossMab, i-DVD Ig, njll. Lawa mapulatifomu axazulula ngempumelelo izinkinga ezifana nokungafani kwe-heavy chain kanye ne-light chain, futhi athuthukise ukufana kanye nokukhiqizwa kwama-antibodies e-bispecific.

Izindlela eziyinhloko zokukhiqizwa kwama-antibody amabili zihlanganisa ukuhlanganiswa kwamakhemikhali, i-hybridoma yomthombo emine, kanye nokulungiswa kwama-antibody obunjiniyela bezakhi zofuzo. Phakathi kwazo, indlela yokuhlanganiswa kwamakhemikhali ihlanganisa izingcezu ezimbili ze-antibody ze-IgG noma ezimbili ze-F (ab ') 2 eziphelele kuma-antibody amabili asebenzisa ama-agent okuhlanganisa amakhemikhali njenge-phthalimide kanye ne-dithioacylbenzoic acid. Le ndlela ilula futhi kulula ukuyisebenzisa, kodwa ingalimaza indawo yokubopha i-antigen, inciphise umsebenzi wama-antibody, futhi i-agent yokuhlanganisa ngokwayo inezinga elithile lokuba nomdlavuza. Indlela ye-hybridoma yomthombo emine isekelwe ekuhlanganisweni kwamaseli e-somatic avela emigqeni emibili ehlukene yamaseli e-hybridoma ukuveza i-IgG yegundane ehambisanayo. Ngobuchwepheshe bobunjiniyela bezakhi zofuzo, ama-antibodies angaguqulwa ngezakhi zofuzo ukuze akhe ama-antibody amabili ahlukile. Ama-antibody amabili ahlukene e-monoclonal akhiwa, futhi izingcezu ze-Fab noma izifunda eziguquguqukayo zeketanga elisindayo kanye neketanga elikhanyayo lama-antibody amabili zahlukaniswa ngokwehlukana. Ngobuchwepheshe bokusabela okuxhumene noma bokuhlukaniswa kweketanga, izingcezu ezimbili zahlanganiswa ukuze zakhe i-antibody amabili ahlukile. Nakuba ubunzima bobuchwepheshe bobunjiniyela bezakhi zofuzo buphezulu kakhulu, okwamanje kuyindlela esetshenziswa kakhulu ekukhiqizweni kwama-antibody amabili ukuze kulungiswe isakhiwo kanye nomsebenzi wama-antibody. Lapho kwenziwa ukwakheka kwama-antibody amabili, isimiso sokusebenza kwama-antibody amabili singasetshenziswa, kodwa ukusabela kwama-antibody amabili kungabangela ukusabela okungekona okuqondile, ngakho-ke kuzocatshangelwa ngokucophelela ekusetshenzisweni okusebenzayo kokwakheka kwama-antibody amabili, njengokwelashwa kwama-antibody amabili.

Ukuhlanzwa kwe-antibody ye-Bispecific inqubo yokuhlukanisa nokuhlanza ama-antibodies e-target ahlanzekile kakhulu. Izindlela ezimbili, i-centrifugation kanye nokuhlunga okujulile, zisetshenziswa ukususa ukungcola okuncibilikayo. I-antibody ye-target ye-bispecific ibanjwa ekuqaleni yi-affinity chromatography. Kuma-antibodies e-IgG afana ne-bispecific, kusetshenziswa i-protein A affinity chromatography, kuyilapho kuma-antibodies angewona i-IgG afana ne-bispecific, i-light chain based affinity chromatography ingasetshenziswa. Ngemva kwalokho, i-antibody ifakwa ngaphansi kwezimo ze-pH ephansi isikhathi esithile, okwenza isakhiwo seprotheni ebusweni be-virus siphazamiseke, ngaleyo ndlela silahlekelwe ikhono lokuthelela amaseli. Ukuhlunga okujulile okuphakathi kwenziwa ukuze kususwe ukungcola okufana nama-protein e-host cell (HCP). Ubumsulwa bama-antibodies buyathuthukiswa ngezindlela ezifana ne-ion exchange chromatography, futhi noma yimaphi amagciwane asele asuswa ngokusebenzisa amasu e-nanofiltration noma i-ultrafiltration. Ekugcineni, isampula igxilisiwe futhi ithathelwe indawo yi-buffer efanele yokwakheka.