Intshayelelo kwiiLayibrari ze-Antibody eziDityanisiweyo

Ilayibrari ye-antibody edibeneyo, eyaziwa ngokuba yilayibrari ye-De novo, yindlela yokwenziwa esebenzisa iindlela ezifana nokuhlanganiswa kwe-DNA okanye ukubonisa i-phage ukuyila nokudibanisa iindawo eziguquguqukayo ze-antibody ezipheleleyo, kuquka iindawo zesakhelo kunye nee-CDR, ngaphandle kokuxhomekeka kwiilayibrari ze-antibody ezingenalwazi.

Ilayibrari ye-antibody e-semi-synthetic yenziwa ngokudibanisa iilayibrari ze-antibody ezenzeka ngokwendalo kunye nokwahluka kokwenziwa. Idla ngokubandakanya ukwenziwa kwee-oligonucleotides ze-DNA ukuvelisa iseti yeendawo ezahlukeneyo ezichazayo ezincedisayo (ii-CDR), ezithi emva koko zidityaniswe nesakhelo se-antibody esingaguqukiyo esithathwe kwimithombo yendalo efana neeseli ze-B zabantu okanye zezilwanyana. I-CDR edibeneyo ingenisa ukwahluka kwilayibrari, nto leyo ekhokelela kwii-antibodies ezijolise kwiindidi ezahlukeneyo zee-epitopes. Ilayibrari ye-antibody e-semi-synthetic inika isivumelwano phakathi kokwahluka kwendalo kwenkqubo yomzimba kunye nokwahluka okulawulwayo okufezekiswa ngeendlela zokwenziwa.

Umahluko phakathi kweelayibrari ze-antibody ezingabonakaliyo kunye nezokwenziwa uxhomekeke kumthombo we-immunoglobulin genes. Okwangoku, umahluko phakathi kwelayibrari ye-antibody eyenziwe kancinci kunye ne-synthesized ulele ekubeni eyangaphambili iqulethe inxalenye yee-antibody segments ezingabonakaliyo, ezifana ne-light chain okanye i-heavy chain, kwaye inye kuphela yazo eyenziwe kwi-vitro; ngelixa leyo yenziweyo ifunyenwe ngokupheleleyo kwi-artificial synthesis yi-PCR in vitro.

I-Alpha Lifetech inokubonelela

I-Alpha Lifetech Inc.inokubonelela ngethala leencwadi le-antibody elenziwe ngokwe-semi-synthesized/synthesized elivela kwizilwanyana nakubantu ngokusekelwe kwiqonga lethu lobuchwephesha lokufumanisa i-antibody. Ngamava eminyaka ekuguqulweni kwezakhi zofuzo kunye nokwakhiwa kwethala leencwadi, amathala eencwadi enziwe ngokwe-synthesized anokwakhiwa anakho ukuvelisa ii-antibodies zokwenziwa ezinomdla kunye neenkcukacha ezingaphaya kwamandla e-antibodies zendalo.I-Alpha Lifetech'sIingcali ziyaziqhenya ngokuvakalisa ukuba singaqinisekisa izinga eliphezulu lempumelelo ekwakhiweni kwethala leencwadi le-antibody eliqulethe ii-clones ezizimeleyo eziyi-10^8 - 10^10.

I-Alpha Lifetech Inc.iyazingca ngokubonelela ngeenkonzo ezipheleleyo zokwakha ithala leencwadi le-antibody ukuhlangabezana neemfuno ezahlukeneyo zabaphandi behlabathi, kubandakanya i-scFv, i-Fab, i-antibody ye-VHH, kunye neelayibrari ezenzelwe wena. Injongo yethu kukuqonda nokuhlangabezana neemfuno zabathengi abahlukeneyo kunye nokunceda ngazo naziphi na iingxaki ezizayo nezivelayo kumsebenzi wophando.

Inkqubo yeNkonzo yoKwakha iiLayibrari ze-Antibody eziDityanisiweyo

Uyilo
Isigaba soyilo siquka ukukhetha ii-antibodies frameworks kunye nemimandla eqinisekisa ukuhambelana (ii-CDR) ngokusekelwe kulandelelwano lwe-antibody ekhoyo okanye idatha yesakhiwo. Ii-antibody ze-CDR ezidibeneyo nazo zinokuyilwa ukwazisa imisebenzi ethile okanye ukuphucula iimpawu zokubopha.

Ukwenziwa kwezinto
I-DNA yokwenziwa ebhala uluhlu lwee-antibody eziyilwe, kuquka zombini iindawo zesakhelo kunye nee-CDR, yenziwa kusetyenziswa iindlela zeekhemikhali okanye ze-enzymatic.

Indibano
Iziqwenga zeDNA ezidityanisiweyo zihlanganiswa zibe ziivektha zokubonakalisa i-antibody kusetyenziswa iindlela ezifana ne-PCR, i-ligation, okanye i-Gibson assembly. Ezi vektha zinokufakwa kwiinkqubo zokubonakalisa ezifana neebhaktheriya, i-yeast, okanye iiseli zezilwanyana ezincancisayo ukuze kuveliswe i-antibody.

Ukuhlola kunye nokukhetha
Iilabhulali zee-antibody ezakhiweyo ziyahlolwa kwaye zikhethwe ii-antibodies ezineempawu ezifunwayo kusetyenziswa iindlela zokuhlola ezisebenzisa i-high-throughput. Oku kunokubandakanya iindlela ezifana ne-phage display, i-yeast display, okanye i-ribosome display, kuxhomekeke kwifomathi yelabhulali kunye nokusetyenziswa kwayo.

Inkonzo Yokwakha Amathala Eencwadi Ezilwa Noku ...

I-Alpha Lifetech Inc.Inika abathengi uyilo olupheleleyo kunye nenkonzo yokwenziwa kweelayibrari ze-antibody ngokudibanisa ii-repertoires ze-V-gene ze-chain ezinzima nezikhaphukhaphu kwi-phage vector isebenzisa inkqubo ye-Cre-lox site-specific recombination ukwenza i-repertoire ye-Fabs eboniswe kwi-phage equka ii-clones eziyi-6.5 × 10^10. Ilayibrari ivelise i-Abs ngokuchasene nee-antigens ezininzi, ezinye ezine-nanomolar affinities. Ukuveliswa kwelayibrari ye-antibody yomntu kusekelwe kwitekhnoloji yokubonisa i-M13 phage (eboniswe kumfanekiso ongezantsi).