Yintoni i-Antibody Engineering?

Ngoncedo lobunjineli be-antibody, kuye kwenzeka ukutshintsha ubungakanani be-molecular, i-pharmacokinetics, i-immunogenicity, i-binding affinity, i-specificity kunye nomsebenzi we-effector wee-antibodies. Emva kokwenza ii-antibodies, ukubopha okuthile kwee-antibodies kuzenza zibe luncedo kakhulu ekuxilongweni kwezonyango nakunyango. Ngobunjineli be-antibody, zinokuhlangabezana neemfuno zophuhliso lwamayeza kunye nokuxilongwa kwangethuba.

Injongo yobunjineli be-antibody kukuyila nokuvelisa imisebenzi ethile ezinzileyo necacileyo engenakufezwa zii-antibodies zendalo, nto leyo ebeka isiseko sokuveliswa kwee-antibodies zonyango.

I-Alpha Lifetech, enamava ayo amakhulu kwiprojekthi yobunjineli be-antibody, inokubonelela ngeenkonzo ze-antibody ze-monoclonal kunye ne-polyclonal ezenzelwe wena kwiintlobo ngeentlobo, kunye neenkonzo zokwakha kunye nokuhlola ithala leencwadi le-antibody ye-phage display. I-Alpha Lifetech inokubonelela abathengi ngee-antibodies ze-biosimilar ezisemgangathweni kunye neemveliso zeproteni eziphinda zihlanganiswe, kunye neenkonzo ezifanayo, ukuvelisa ii-antibodies ezisebenzayo, ezichanekileyo kakhulu, nezizinzileyo. Ngokusebenzisa ii-antibody ezipheleleyo, amaqonga eproteni kunye neenkqubo zokubonisa i-phage, sinikezela ngeenkonzo ezigubungela imveliso ye-antibody ephezulu nesezantsi, kubandakanya iinkonzo zobugcisa ezifana nokwenziwa kwe-antibody humanization, ukucocwa kwe-antibody, ukulandelelana kwe-antibody, kunye nokuqinisekiswa kwe-antibody.

Inqanaba lokuqala lobunjineli be-antibody linxulumene neetekhnoloji ezimbini:

--Itekhnoloji ye-DNA ephinda-phindayo

--Iteknoloji yeHybridoma

Uphuhliso olukhawulezileyo lobunjineli be-antibody lunxulumene neetekhnoloji ezintathu ezibalulekileyo:

--Iteknoloji yokudibanisa iijini kunye ne-polymerase chain reaction

--Ukubonakaliswa kweproteni: Iiproteni eziphinda-phindayo ziveliswa ziinkqubo zokubonakaliswa ezifana nemvubelo, iintsholongwane ezimile okwentonga, kunye nezityalo

--Uyilo lwesakhiwo oluxhaswa yikhompyutha

Isizukulwana sokuqala see-antibodies senziwe saba ngabantu ukuze kuveliswe ii-antibodies ze-chimeric, apho indawo eguquguqukayo yee-antibodies ze-monoclonal zegundane yayidityaniswe nendawo ehlala ihleli yee-molecule ze-IgG zabantu. Indawo yokubopha i-antigen (CDR) ye-antibody ye-monoclonal yesizukulwana sesibini segundane yatyalwa kwi-IgG yabantu. Ngaphandle kwengingqi ye-CDR, zonke ezinye ii-antibodies ziphantse zibe zii-antibodies zabantu, kwaye kwenziwa imizamo yokuphepha ukuphembelela iimpendulo ze-antibody yegundane (HAMA) yabantu xa kusetyenziswa ii-antibodies ze-clone zegundane kunyango lwabantu.

 

Ukwakha ithala leencwadi lokubonisa i-phage, inyathelo lokuqala kukufumana ii-genes ezifaka ii-antibodies, ezinokuhlukaniswa kwiiseli ze-B zezilwanyana ezigonyiweyo (ukwakhiwa kwethala leencwadi lokuzikhusela), ezikhutshwe ngqo kwizilwanyana ezingagonywanga (ukwakhiwa kwethala leencwadi lendalo), okanye zihlanganiswe kwi-vitro kunye neziqwenga ze-antibody gene (ukwakhiwa kwethala leencwadi elenziweyo). Emva koko, ii-genes ziyandiswa yi-PCR, zifakwe kwii-plasmids, kwaye zibonakaliswe kwiinkqubo ezifanelekileyo zomninimzi (ukubonakaliswa kwe-yeast (ngesiqhelo iPichia pastoris), ukubonakaliswa kwe-prokaryotic (ngesiqhelo i-E. coli), ukubonakaliswa kweeseli zezilwanyana ezincancisayo, ukubonakaliswa kweeseli zezityalo, kunye nokubonakaliswa kweeseli zezinambuzane ezisuleleke ziintsholongwane ezimile njengentonga). Eyona ixhaphakileyo yinkqubo yokubonakaliswa kwe-E. coli, edibanisa ulandelelwano oluthile lwe-encoding antibody kwi-phage kwaye ifaka enye yeeproteni ze-phage shell (pIII okanye pVIII). Ukuhlanganiswa kwe-gene ye, Kwaye iboniswe kumphezulu we-bacteriophages. Ingundoqo yale teknoloji kukwakha ithala leencwadi lokubonisa i-phage, elinenzuzo ngaphezu kweelayibrari zendalo kuba linokuba nokubopha okuthile. Emva koko, ii-antibodies ezine-antigen specificity ziyahlolwa ngenkqubo yokukhetha izinto eziphilayo, ii-antigens ezijoliswe kuzo ziyalungiswa, ii-phages ezingabotshwanga ziyahlanjwa ngokuphindaphindiweyo, kwaye ii-phages ezibotshiweyo ziyahlanjwa ukuze ziphuculwe ngakumbi. Emva kokuphindaphinda izihlandlo ezithathu nangaphezulu, ii-antibodies ezine-specificity ephezulu kunye ne-high affinity ziyahlulwa.

Umfanekiso 3: Ukwakhiwa kunye nokuhlolwa kweThala leeNcwadi le-Antibody

Itekhnoloji ye-Recombinant DNA ingasetyenziselwa ukuvelisa iziqwenga ze-antibody. Ii-antibodies ze-Fab ekuqaleni zinokufakwa kuphela kwi-gastric protease ukuvelisa iziqwenga ezi-2 (Fab '), ezithi emva koko zigaywe yi-papain ukuvelisa iziqwenga ze-Fab nganye. Isiqwenga se-Fv siquka i-VH kunye ne-VL, ezizinzileyo ngenxa yokungabikho kweebhondi ze-disulfide. Ke ngoko, i-VH kunye ne-VL zidityaniswa kunye nge-peptide emfutshane ye-15-20 amino acids ukwenza i-antibody ye-single chain variable fragment (scFv) enobunzima be-molecular obumalunga ne-25KDa.

Uphononongo lwesakhiwo se-antibody kwiCamelidae (Camel, LIama, kunye neAlpaca) lucacisile ukuba ii-antibodies zinemixokelelwane enzima kuphela kwaye azinamixokelelwane ekhanyayo, yiyo loo nto zibizwa ngokuba zii-heavy chain antibodies (hcAb). Indawo eguquguqukayo yee-heavy chain antibodies ibizwa ngokuba yi-single domain antibodies okanye ii-nanobodies okanye i-VHH, ezinobukhulu obuyi-12-15 kDa. Njengee-monomers, azinamixokelelwane ye-disulfide kwaye zizinzile kakhulu, zinobuhlobo obuphezulu kakhulu kwii-antigens.

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

Ukubonakaliswa kwe-cell free expression kusebenzisa ukubonakaliswa kwe-DNA yendalo okanye yokwenziwa ukuze kufezekiswe ukwenziwa kwe-in vitro protein, ngokuqhelekileyo kusetyenziswa inkqubo yokubonakaliswa kwe-E. coli. Ivelisa iiproteni ngokukhawuleza kwaye iphephe umthwalo we-metabolic kunye ne-cytotoxic kwiiseli xa ivelisa inani elikhulu leeproteni eziphinda-phindayo emzimbeni. Ingavelisa neeproteni ezinzima ukuzivelisa, njengezo kunzima ukuziguqula emva kokuguqulwa okanye ukwenza iiproteni ze-membrane.