Isingeniso Semitapo Yolwazi Yamasosha Ahlanganisiwe

Ilabhulali ye-antibody ehlanganisiwe, eyaziwa nangokuthi umtapo wezincwadi we-De novo, iyindlela yokwenziwa esebenzisa amasu afana nokuhlanganiswa kwe-DNA noma isibonisi se-phage ukuze udizayine futhi uhlanganise izifunda eziphelele eziguquguqukayo ze-antibody, okuhlanganisa izifunda zohlaka kanye nama-CDR, ngaphandle kokuthembela kumalabhulali e-naïve antibody.

Ilabhulali ye-semi-synthetic antibody idalwe ngokuhlanganisa imitapo yolwazi eyenzeka ngokwemvelo nokuhlukahluka kokwenziwa. Ngokuvamile kuhilela ukuhlanganisa ama-oligonucleotide e-DNA ukuze kukhiqizwe isethi yezifunda ezinqumayo ezihambisanayo (CDRs), ezihlanganiswa nohlaka oluqinile lwe-antibody olutholakala emithonjeni yemvelo njengamaseli B omuntu noma esilwane. I-CDR ehlanganisiwe yethula ukwehlukahlukana emtatsheni wezincwadi, okuholela ekutheni amasosha omzimba aqondise izinhlobonhlobo zama-epitopes. Umtapo wolwazi we-semi-synthetic antibody uhlinzeka ngokuvumelana phakathi kokwehlukahlukana kwemvelo kwamasosha omzimba kanye nokwehlukahlukana okulawulwayo okuzuzwa ngezindlela zokwenziwa.

Umehluko phakathi kwemitapo yolwazi ye-naïve neye-synthetic antibody uncike emthonjeni wezakhi zofuzo ze-immunoglobulin. Khonamanjalo, umehluko phakathi komtapo wolwazi wamasosha omzimba ohlanganiswe kancane futhi ohlanganisiwe usekutheni owokuqala uqukethe ingxenye yamasegmenti e-naïve antibody, njengeketango elilula noma iketango elisindayo, futhi eyodwa kuphela yawo ehlanganiswa nge-vitro; kuyilapho ehlanganisiwe isuselwa ngokuphelele ekuhlanganiseni okwenziwe yi-PCR in vitro.

I-Alpha Lifetech ingahlinzeka

Inkampani Alpha Lifetech Inc.inganikeza umtapo wolwazi wamasosha omzimba ohlanganiswe kancane/ohlanganisiwe ovela ezilwaneni nakubantu ngokusekelwe kungxenyekazi yethu yochwepheshe yokutholwa kwamasosha omzimba. Ngesipiliyoni seminyaka ekuguqulweni kofuzo kanye nokwakhiwa komtapo wolwazi, imitapo yolwazi edidiyelwe ingakhiwa ekwazi ukukhiqiza amasosha omzimba okwenziwa ahambisana nokucaciswa okungaphezu kwamandla amasosha omzimba emvelo.I-Alpha Lifetech'sochwepheshe bayaziqhenya ngokumemezela ukuthi singaqinisekisa izinga eliphezulu lempumelelo ekwakhiweni komtapo wolwazi wamasosha omzimba aqukethe 10^8 – 10^10 ama-clone azimele.

Inkampani Alpha Lifetech Inc.iyaziqhenya ngokunikela ngezinsiza ezibanzi zokwakha umtapo wolwazi we-antibody ukuhlangabezana nezidingo ezahlukahlukene zabacwaningi bomhlaba wonke, okuhlanganisa i-scFv, i-Fab, i-VHH antibody, nemitapo yolwazi eyenziwe ngokwezifiso. Inhloso yethu ukuqonda kanye nokuhlangabezana nezidingo zamakhasimende ahlukene kanye nokusiza kunoma yiziphi izinkinga ezizayo nezivelayo emsebenzini wocwaningo.

Inqubo Yesevisi Yokwakhiwa Kwemitapo Yolwazi Ehlanganisiwe Ye-antibody

Idizayini
Isigaba sokuklama sibandakanya ukukhetha izinhlaka ze-antibody kanye nezifunda ezinquma ukuhambisana (CDRs) ngokusekelwe ekulandeleni okukhona kwama-antibody noma idatha yesakhiwo. Amasosha omzimba okwenziwa e-CDR angaphinda adizayinelwe ukwethula imisebenzi ethile noma athuthukise izakhiwo ezibophayo.

I-synthesis
I-synthetic DNA efaka amakhodi ama-antibody aklanyelwe ukulandelana, okuhlanganisa kokubili izifunda zohlaka kanye nama-CDR, ahlanganiswa kusetshenziswa izindlela zamakhemikhali noma ze-enzymatic.

Umhlangano
Izingcezu ze-DNA ezihlanganisiwe zihlanganiswa zibe ama-antibody expression vectors kusetshenziswa amasu afana ne-PCR, ligation, noma i-Gibson assembly. Lawa ma-vector angabe esengeniswa ezinhlelweni zokubonisa ezifana nebhaktheriya, imvubelo, noma amangqamuzana ancelisayo ukuze kukhiqizwe amasosha omzimba.

Ukuhlolwa kanye Nokukhetha
Imitapo yolwazi eyakhiwe yama-antibody iyahlolwa futhi ikhethelwa amasosha omzimba anezakhiwo ezifiselekayo kusetshenziswa izindlela zokuhlola eziphezulu. Lokhu kungase kuhlanganise amasu anjengombukiso we-phage, isibonisi seyeast, noma isibonisi se-ribosome, kuye ngefomethi yelabhulali nokusebenza.

Isevisi Yokwakhiwa Kwemitapo Yama-antibody Ehlanganisiwe

Inkampani Alpha Lifetech Inc.inikeza amakhasimende umklamo wokuma okukodwa kanye nensizakalo ehlanganisiwe yemitapo yolwazi yamasosha omzimba ngokuhlanganisa uchungechunge olusindayo nolulula lwe-V-gene repertoires ku-vector ye-phage kusetshenziswa isistimu ye-Cre-lox yesayithi yokuhlanganisa kabusha ukuze yakhe iqoqo le-Fabs ekhonjiswe ku-phage ehlanganisa ama-clone angu-6.5 × 10^10. Umtapo wezincwadi ukhiqize i-Abs ngokumelene nama-antigen amaningi, amanye ane-nanomolar affinities. Ukwenziwa komtapo wolwazi wamasosha omzimba kusekelwe kubuchwepheshe bokubonisa i-M13 phage (eboniswe emfanekisweni ongezansi).