Iqonga loPhuhliso lwee-Antibody eziBispecific

I-Alpha Lifetech inokubonelela abathengi ngokufumanisa i-antibody esemgangathweni oqinisekisiweyo kunye neemveliso kunye neenkonzo zeproteni ephinda-phindayo. Singalungiselela ii-antibodies ngokusebenza kakuhle, ukucacisa okuqinileyo, kunye nozinzo oluhle. I-Alpha Lifetech inezixhobo ezahlukeneyo zokucoca ii-antibody kunye nezixhobo, ezinokubonelela ngeenkonzo zokucoca ii-antibody ezivela kwimithombo eyahlukeneyo efana nee-antibodies zomvundla, igusha, inkukhu, kunye neempuku, kunye neenkonzo zokucoca i-affinity yeProtein A/G kunye neenkonzo zokwahlula kunye nokucoca ii-antibody. Ngokusekelwe kulwakhiwo olupheleleyo lwenkqubo yeqonga lokufumanisa ii-antibody, iqonga leproteni, njl.njl., sigubungela iinkonzo zokuvelisa ii-antibody ezisezantsi neziphezulu, kwaye sinokubonelela ngeenkonzo zobugcisa ezivela kulungiselelo lwee-antibody, ukuhlanjululwa kwee-antibody ezimbini kunye nokwahlula kunye nokucoca ii-antibody, ukulandelelana kwee-antibody, ukuqinisekiswa kwee-antibody, njl.njl., ezinokusetyenziselwa unyango lwee-antibody ezimbini.

Ii-antibodies ze-Monoclonal zibhekisa kwi-immunoglobulins eziveliswa yiseli enye ye-B eneenkcukacha eziphezulu zee-antigens okanye ii-epitopes. Ukuveliswa kwee-antibodies ze-Monoclonal kwaqalwa kwalungiselelwa kusetyenziswa iteknoloji ye-hybridoma ukuvelisa ii-antibodies zegundane. Ngokukodwa, ngokudibanisa iiseli ze-spleen ezivela kwiimpuku ezigonyiweyo kunye neeseli ze-myeloma ezivela ebantwini okanye kwiimpuku, kwenziwa iiseli ze-hybridoma, ezikhupha ii-antibodies ezithile. Ii-antibodies ze-monoclonal eziveliswayo zempuku zisetyenziswa kakhulu kuphando lwezilwanyana kunye nokuxilongwa kwezifo emva kweenkonzo zokucoca. Nangona kunjalo, umzimba womntu uya kuvelisa impendulo yomzimba kwiiproteni zempuku zangaphandle, ngoko ke kukho imida ebalulekileyo ekusetyenzisweni kweklinikhi kokuveliswa kwee-antibodies ze-monoclonal zempuku. Kamva, abantu basebenzisa "ukwenziwa komntu" ukulungiselela ii-antibodies ze-monoclonal ezivela kwiimpuku, kwaye basebenzisa iteknoloji yobunjineli be-genetic ukuguqula ii-antibodies zempuku ukuba zibe nommandla oqhubekayo we-immunoglobulin yomntu ukunciphisa ukuqina kwazo kwe-immunogenicity. Olu hlobo lwe-antibody lukwaziwa njenge-antibody ye-monoclonal yomntu. Ukongeza, ukusebenzisa iiseli zabantu ukuvelisa ii-antibodies ze-monoclonal kubizwa ngokuba kukulungiselela i-antibody ye-monoclonal yomntu wonke. Ii-antibodies ze-Monoclonal zisetyenziswa kakhulu kwizicelo ze-biomedical kunye ne-clinical. Kuphando lwezonyango, ii-antibodies ze-monoclonal zingasetyenziswa njengee-immunomodulators, ii-antibodies ze-monoclonal zonyango zingasetyenziswa kwi-chemotherapy yomhlaza kunye nonyango losulelo lwentsholongwane, kwaye uphuhliso lwee-conjugates ze-antibody ze-monoclonal lusetyenziswa kakhulu kunyango lomhlaza.

Intshayelelo kwi-Bispecific Antibody

Ngo-1960, kwacetyiswa ingcamango yee-antibodies ze-bispecific. Ii-antibodies ze-bispecific, ezaziwa ngokuba zii-antibodies ze-bispecific monoclonal, zii-antibodies ezenziwe ngokwenziwa kusetyenziswa iteknoloji yobunjineli be-genetic. Njenge-antibody yokwenziwa eyenziweyo, ii-antibodies ze-bispecific zihlala ziyinxalenye ye-IgG kwaye ziqulathe iqhekeza le-antigen elijolise kwi-subunit ye-CD3. Ii-antibodies ze-bispecific zineendawo ezimbini ezithile zokubopha i-antigen ezinokubopha kwaye ziqaphele ii-antigen ezimbini ezahlukeneyo ngaxeshanye, okanye ii-epitopes ezimbini ezahlukeneyo ze-antigen. Xa kuthelekiswa nee-antibodies ze-monoclonal, ii-antibodies ze-bispecific zinendawo eyongezelelweyo yokubopha i-antigen, ngaloo ndlela zinamandla okucacisa kunye nokukwazi ukujolisa, ezinokujolisa ngokuchanekileyo iiseli ze-tumor kwaye zinciphise ubuthi obungajoliswanga. Ii-antibodies ze-bispecific zinokwenza imisebenzi emininzi yebhayoloji ngaxeshanye, njengokuqokelela iiseli zomzimba, ukuvala iindlela zokubonisa, kunye nokubulala ngokuthe ngqo iiseli ze-tumor. Ii-antibodies zokuqala ze-bispecific zazilungiselelwe ngokudityaniswa kweekhemikhali okanye ukuhlanganiswa kweeseli, kodwa le ndlela isenokuba ihambe kancinci ngenxa yokudibana okungacwangciswanga kunye nobunzima bokuhlukanisa indibaniselwano ekujoliswe kuyo. Ngophuhliso oluqhubekayo lobuchwepheshe bobunjineli bemfuza, kuye kwaphuhliswa amaqonga amaninzi amatsha eteknoloji, afana namaqhina kwimingxunya (KIH), iCrossMab, iDVD Ig, njl. La maqonga asombulula ngempumelelo iingxaki ezifana nokungahambelani kweetsheyini ezinzima kunye neetsheyini ezikhanyayo, kwaye aphucula ukufana kunye nokuveliswa kwee-antibodies ze-bispecific.

Umzobo 1 Isishwankathelo esilula sesicwangciso seendlela ezicetywayo zokusebenza kwee-antibodies ze-bispecific (bsAbs) kwizilingo zeklinikhi ze-oncology.(Umfanekiso womthombo: Uphononongo lwee-antibodies ze-bispecific kunye nezakhiwo ze-antibody kwi-oncology kunye nemingeni yeklinikhi - ScienceDirect)

Itekhnoloji yokuvelisa ii-Antibodies ezimbini

Iindlela eziphambili zokuvelisa i-antibody ye-bispecific ziquka ukuhlanganiswa kweekhemikhali, i-hybridoma yemvelaphi emine, kunye nokulungiswa kwe-antibody yobunjineli be-genetic. Phakathi kwazo, indlela yokudibanisa iikhemikhali idibanisa iziqwenga ezimbini ze-antibody ze-IgG okanye ezimbini ze-F (ab ') 2 ezingasebenziyo kwi-antibody ye-bispecific isebenzisa ii-agents ze-chemical coupling ezifana ne-phthalimide kunye ne-dithioacylbenzoic acid. Le ndlela ilula kwaye kulula ukuyisebenzisa, kodwa inokonakalisa indawo yokubopha i-antigen, inciphise umsebenzi we-antibody, kwaye i-agent yokudibanisa ngokwayo inomlinganiselo othile we-carcinogenicity. Indlela ye-hybridoma yemvelaphi emine isekelwe ekudibaneni kweeseli ze-somatic ezivela kwimigca emibini eyahlukeneyo yeeseli ze-hybridoma ukuze kuvezwe i-IgG yegundane ehambelanayo. Ngeteknoloji yobunjineli be-genetic, ii-antibodies zinokuguqulwa ngokwe-genetic ukuze zenze ii-antibodies ze-bispecific. Ii-antibodies ezimbini ezahlukeneyo ze-monoclonal zakhiwa, kwaye iziqwenga ze-Fab okanye iindawo eziguquguqukayo ze-chain heavy kunye ne-light chain zamaqela amabini e-antibodies zahlulwa ngokwahlukeneyo. Ngeteknoloji yokudibanisa okanye yokudibanisa i-chain, ezi ziqwenga zimbini zadityaniswa ukuze zenze i-antibody ye-bispecific. Nangona ubunzima bobuchwephesha bobunjineli bemfuza buphezulu kakhulu, okwangoku yeyona ndlela isetyenziswa kakhulu kwimveliso ye-bispecific antibody ukulungisa ulwakhiwo kunye nomsebenzi we-antibodies. Xa kusenziwa uyilo lwe-bispecific antibody, umgaqo we-antibody cross reactivity ungasetyenziswa, kodwa i-antibody cross reactivity inokubangela iimpendulo ezingezizo ezithile, ngoko ke iya kuqwalaselwa ngononophelo ekusebenziseni uyilo lwe-bispecific antibody, njengonyango lwe-bispecific antibody.

Ukucocwa kwee-Antibodies zeBispecific

Ukucocwa kwe-antibody ye-Bispecific yinkqubo yokwahlula nokucoca ii-antibodies ezijoliswe kwi-high-purity target. Iindlela ezimbini, i-centrifugation kunye ne-deep filtration, zisetyenziselwa ukususa ezinye izinto ezingcolisayo. I-antibody ye-target bispecific ibanjwa kuqala yi-affinity chromatography. Kwi-antibodies ze-IgG ezifana ne-bispecific, kusetyenziswa i-protein A affinity chromatography, ngelixa kwi-non-IgG ezifana ne-bispecific antibodies, i-light chain based affinity chromatography ingasetyenziswa. Emva koko, i-antibody ifakwa phantsi kweemeko ze-pH ephantsi kangangexesha elithile, nto leyo ebangela ukuba isakhiwo seproteni kumphezulu we-virus siphazamiseke, ngaloo ndlela silahlekelwe ngamandla okosulela iiseli. Ukuhluzwa okunzulu okuphakathi kwenziwa ukuze kususwe izinto ezingcolisayo ezifana neeproteni ze-host cell (HCP). Ubumsulwa bee-antibodies buphuculwa ngeendlela ezifana ne-ion exchange chromatography, kwaye naziphi na iintsholongwane ezisele zisuswa ngokusebenzisa iindlela ze-nanofiltration okanye ze-ultrafiltration. Ekugqibeleni, isampuli ixutyiwe kwaye ithathelwe indawo yi-formulation buffer efanelekileyo.

Ukuhamba kweNkonzo yoMveliso wee-Antibodies zeBispecific

Amanyathelo	Umxholo weNkonzo	Umgca wexesha
Ukwenziwa kweGene	Ukuveliswa kwezicwangciso zoyilo ezahlukeneyo ezifikelela kwi-3, ukuveliswa kwe-DNA plasmids ye-recombinant antibody	Iiveki ezi-2-3
Uvavanyo oluNcinci	Ukuveliswa kweeproteni ze-antibody ezimbini → Ukubonakaliswa okuncinci kwimigca yeeseli zezilwanyana ezincancisayo → Ukuqinisekiswa yi-SDS-PAGE → Uhlalutyo lokubopha ngee-antigen zeproteni eziphinda zibunjwe yi-ELISA → Uthelekiso lwee-clones kunye nee-antibodies zabazali	Iiveki ezi-5-6
Chonga	Ukubonakaliswa kwee-antibodies ezimbini zokuqala ezipheleleyo ze-bispecific → Uhlalutyo lokubopha yi-ELISA → Uvavanyo lwe-ELISA lwee-antibodies eziqatshelwe zii-antibodies ze-anti mouse	Iveki e-1
Ukuveliswa kwee-Antibody	Ukuveliswa kwee-antibody ezinkulu	Iiveki ezi-3-4

Iingenelo Zethu

Sigubungela iinkonzo zokuvelisa ii-antibody ezisezantsi neziphezulu, kwaye sinokubonelela ngeenkonzo zobugcisa ukusuka ekulungiseleleni ii-antibody, ukucocwa kwee-antibody ezimbini kunye nokwahlulwa kunye nokuhlanjululwa kwee-antibody, ukulandelelana kwee-antibody, ukuqinisekiswa kwee-antibody, njl.njl., ezinokusetyenziselwa unyango lwee-antibody ezimbini.

Ukuphakama Okuphezulu

Ukubonakaliswa kweprotheyini ephezulu kwiiseli ze-CHO.
Indawo yokugcina ezinzileyo

Iiveki ezingaphezu kwezi-2 kwi-serum kwi-37°C.
Akukho Ngxaki Yokunyibilika

>30 mg/ml
Iiseli eziZinzileyo

Iiseli ezininzi ezizinzileyo ezinee-antigen ze-tumor ziye zaphuhliswa ukuqinisekisa ii-antibodies ezininzi ze-bispecific