I-Phage Display Antibody Development Platform

I-Alpha Lifetech, enokuhlangenwe nakho kwayo okubanzi kwephrojekthi ekutholeni amasosha omzimba, ingahlinzeka ngezinsizakalo ezenziwe ngokwezifiso ze-monoclonal kanye ne-polyclonal antibody yezinhlobo eziningi, kanye nokwakhiwa komtapo wolwazi wamasosha omzimba kanye nezinsizakalo zokuhlola. I-Alpha Lifetech inganikeza amakhasimende amasosha omzimba aqinisekisiwe ikhwalithi kanye nemikhiqizo yamasosha omzimba aphinde ahlanganiswe, kanye nezinsizakalo ezihambisanayo, ukuze alungiselele amasosha omzimba asebenza kahle, acace kakhulu, futhi azinzile. Ngokusebenzisa izinkundla eziphelele zokulwa nama-antibody nobunjiniyela bamasosha omzimba, sihlinzeka ngezinsizakalo ezimboza umfula okhuphukayo kanye nophansi kokukhiqizwa kwamasosha omzimba, okuhlanganisa nezinsizakalo zobuchwepheshe ezifana nokulungiselela amasosha omzimba, ukuhlanzwa, ukulandelana kwe-antibody, nokuqinisekisa.

I-Alpha Lifetech inenkundla yokuthola i-antibody ekhulile ehlinzeka amakhasimende ngezinsiza eziningi ezihlanganisa ukungavikeleki kwezilwane, ukwakhiwa nokuhlolwa komtapo wezincwadi we-phage, ukulandelana kwe-antibody, ukuvela kwe-antibody, ukuhlanzwa kwe-antibody, ukuqinisekiswa kwe-antibody, kanye nokulebula kwe-antibody okusekelwe kubuchwepheshe be-hybridoma, ubuchwepheshe be-cell B eyodwa, ubuchwepheshe bokubonisa i-phage, nokuningi. I-Alpha Lifetech inolayini wokukhiqiza ojwayelekile, futhi ngaphezu kwezinsizakalo zokulungiselela ama-antibody, izinsizakalo ezisekelayo ezifana nokwenza abantu bamasosha omzimba, ukuvuthwa kwe-antibody affinity, ukwakhiwa nokuthuthukiswa kwezidakamizwa ze-ADC, kanye nomklamo wokulandelana olandelanayo we-CAR-T nakho kunganikezwa. Ngesikhathi esifanayo, i-Alpha Lifetech yakhe umtapo wolwazi ovikela amasosha omzimba ngokusekelwe ku-M13, T4, ne-T7 phage yesikhulumi sobuchwepheshe, esinamandla okugcina afinyelela ku-10^8-10^9. Izinga elihle, izinga lokufaka, kanye nokwehlukahlukana komtapo wolwazi konke kungafinyelela ngaphezu kwama-90%.

I-Phage display Technology

Ukukhiqizwa kwamasosha omzimba e-Monoclonal ekuqaleni kwalungiselelwa kusetshenziswa ubuchwepheshe be-hybridoma monoclonal antibody ukukhiqiza igundane le-hybridoma monoclonal antibody. Ngokukhethekile, ngokuhlanganisa amangqamuzana ombende kumagundane agonyiwe namaseli e-myeloma avela kubantu noma amagundane, kwakheka amaseli e-hybridoma, akhiqiza amasosha omzimba e-hybridoma monoclonal antibody. Ngokuguqulwa komuntu kwamasosha omzimba egundane kanye nokuguqulwa kobunjiniyela bofuzo kwamasosha omzimba ukuze abanike indawo ehlala njalo ye-immunoglobulin yomuntu, ukuzivikela kwawo kungancishiswa. Ubunjiniyela be-Monoclonal antibody busetshenziswa kabanzi ezinhlelweni ze-biomedical nezomtholampilo.

I-Alpha Lifetech ingahlinzeka ngezinsizakalo zobunjiniyela be-antibody eyodwa-stop kusukela ekwakhiweni kwephrothokholi yocwaningo lwangaphambi kwezempilo kuye ekutholweni kwe-antibody drug conjugates (ADCs) kanye nokuqinisekiswa kwezilwane. Ama-antibody drug conjugates (ADCs) angaletha imishanguzo ye-chemotherapy kumaseli omdlavuza. Ngemuva kokubophezela ekuhlosweni okuqondile okuvezwe kumaseli omdlavuza, i-ADC ikhipha izidakamizwa eziyi-cytotoxic kumaseli omdlavuza. I-Alpha Lifetech ingase futhi inikeze amakhasimende izinsiza zokuvuthwa kwe-antibody affinity. Ngokuthuthukiswa kokuguquguquka okuthuthukisiwe kanye nobuchwepheshe bokuhlola isibonisi se-phage esiphezulu, amasosha omzimba anobudlelwane obuthile ayahlolwa kuqala, bese kwethulwa ukuguqulwa kwama-amino acid ukuze kukhiqizwe okuhlukile. Kamuva, amasosha omzimba asondelene kakhulu ahlolwa futhi ahlolwa kusetshenziswa amasu okuhlola. Ngemuva kwemizuliswano eminingi yokwenza kahle kanye nokuhlaziywa kwesakhiwo, amasosha omzimba anobungane obuphezulu kanye nokucaciswa okuqinile kwatholakala ekugcineni.

I-Alpha Lifetech ingakha izinhlobo ezahlukene zamalabhulali wokubonisa i-antibody phage, okuhlanganisa imitapo yolwazi yokuzivikela komzimba, imitapo yolwazi yomdabu, imitapo yolwazi yokwenziwa engalingani, nemitapo yolwazi yokwenziwa. Ngokusekelwe kumthamo ophezulu wemitapo yolwazi yamasosha omzimba, amasosha omzimba akhethekile angatholwa. Amavektha amaningi we-phagemid afana ne-pMECS, pComb3X, ne-pCANTAB 5E anganikezwa, kanye nezinhlobo ezifana ne-TG1 E. coli, XL1 Blue, ne-ER2738. Ngaphezu komthamo womtapo wolwazi ofinyelela ku-10^9, izinga lokufakwa kweziqephu ezihlosiwe zomtapo wolwazi nalo liphezulu, okudala izimo ezanele zokuhlola amasosha omzimba anelisa amakhasimende. I-flowchart yesevisi yokulungiselela amasosha omzimba ye-Alpha Lifetech esekelwe kubuchwepheshe bokubonisa ikheji iboniswa kuMfanekiso 1.

Umdwebo we-Fig.1 Umdwebo wenqubo yokulungiselela amasosha omzimba we-monoclonal ngokusekelwe kubuchwepheshe bokubonisa i-phage.

I-Phage Display Antibody Production Workflow

Izinyathelo	Okuqukethwe Kwesevisi	Umugqa wesikhathi
Isinyathelo 1: Ukugonywa kwezilwane	(1) Ukugonywa kwezilwane izikhathi ezi-4, ukugonywa kwe-booster 1 umthamo, isamba semithamo emi-5 egonyiwe. (2) I-serum engemihle ngaphambi kokuba kuqoqwe ukugonywa, futhi i-ELISA yenziwa ngethamo lesine ukuthola i-serum titer. (3) Uma i-serum antibody titer yomthamo wesine ihlangabezana nezimfuneko, umthamo owodwa owengeziwe wokugoma uzonikezwa izinsuku ezingu-7 ngaphambi kokuqoqwa kwegazi. Uma ingahlangabezani nezidingo, ukugoma okujwayelekile kuzoqhubeka. (4) Amandla afanelekayo, ukuqoqwa kwegazi nokuhlukaniswa kwama-monocyte	10 amasonto
Isinyathelo sesi-2: Ukulungiselela i-cDNA	(1) I-PBMC Ingqikithi Ye-RNA Extraction (Ikhithi Yokudonsa ye-RNA) (2) Ukulungiswa okuphezulu kwe-RT-PCR ye-cDNA (ikhithi yokuhlehla yokubhala)	1 usuku
Isinyathelo sesi-3: Ukwakhiwa Kwelabhulali Ye-Antibody	(1) Ngokusebenzisa i-cDNA njengesifanekiso, izakhi zofuzo zandiswa imizuliswano emibili ye-PCR. (2) Ukwakhiwa kwe-Phage nokuguqulwa: i-gene splicing phagemid vector, ukuguqulwa kwe-electroporation ye-TG1 host bacteria, ukwakhiwa komtapo wolwazi we-antibody. (3) Ukuhlonza: Khetha ngokungahleliwe ama-clones angu-24, ukuhlonza i-PCR isilinganiso esihle+izinga lokufaka. (4) Ukulungiswa kwephage okusizwayo: I-M13 phage amplification + purification. (5) Ukuhlenga umtapo wolwazi wephage	3-4 amasonto
Isinyathelo sesi-4: Ukuhlolwa Kwelabhulali Ye-Antibody (imizuliswano emi-3)	(1) Ukuhlola okuzenzakalelayo okuyindilinga engu-3 (ukuhlolwa kwesigaba esiqinile): ukuhlolwa kwengcindezi ukuze kukhishwe amasosha omzimba angaqondile ngokwezinga eliphezulu ngangokunokwenzeka. (2) I-bacteriophage yokukhulisa i-clone eyodwa ekhethiwe+I-IPTG induced expression+ELISA ukutholwa kwama-clones amahle. (3) Wonke ama-clone amahle akhethelwe ukulandelana kwezakhi zofuzo.	4-5 amasonto

Inzuzo Yobuchwepheshe Bokubonisa Iphage

I-Alpha Lifetech inokuhlangenwe nakho okucebile emkhakheni wokuthuthukiswa kwamasosha omzimba. Ngokuhamba kweminyaka, i-Alpha Lifetech isungule inkundla yokuthuthukisa ama-antibody.

Izinsizakalo Zokusekela

Singahlinzeka ngezinsizakalo ezahlukene zokwakha umtapo wolwazi wokuzivikela ezifweni kanye nezinsizakalo zokuhlola umtapo wolwazi wemvelo we-antibody ngokuya ngezidingo zamakhasimende

Okuhlosiwe Okuningi

Izinsiza eziningi zokutholwa kwe-antibody okuhlosiwe ziyatholakala: amaprotheni, ama-peptide, ama-molecule amancane, amagciwane, amaprotheni e-membrane, i-mRNA, njll.

Ama-Vector amaningi

Isevisi yokwakha umtapo wolwazi womuntu siqu, singahlinzeka ngama-bacteriophage vectors ahlukahlukene ahlanganisa i-PMECS, i-pComb3X, ne-pCANTAB 5E, futhi sizilungise ngokwezidingo zamakhasimende.

Inkundla Evuthiwe

Umthamo wesitoreji ungafinyelela ku-10 ^ 8-10 ^ 9, amanani okufaka wonke angaphezulu kwama-90%, futhi ukuhambisana kwamasosha omzimba atholakala ngokuhlolwa ngokuvamile kusezingeni le-nM pM.

INKONZO Ehlobene

Amasu Okuthuthukisa Amasosha Ahlukahlukene

Isevisi Yokuthuthukisa I-Monoclonal Antibody

I-Hybridoma Technology Platform

I-Single B Cell Sotting Platform

I-Phage Display Antibody Development Platform

ukuthuthukiswa kwe-monoclonal antibody-Alpha Lifetech

Isevisi Yokuthuthukisa I-Monoclonal Antibody

Singahlinzeka ngezinsiza zekhwalithi ephezulu, ezimsulwa, nezicaciswe kakhulu zokuthuthukisa i-monoclonal antibody, okuhlanganisa ukukhiqizwa kwamasosha omzimba egundane kanye namasosha omzimba kanogwaja.

I-Hybridoma Technology Platform

Kubandakanya uhlelo lokugoma, izinsiza zokulungiselela amasosha omzimba, ukuhlanzwa kwe-antibody, ukulandelana kokusebenza okuphezulu kwe-antibody, ukuqinisekiswa kwe-antibody, njll.

I-Single B Cell Sotting Platform

I-Alpha Lifetech inezinzuzo ngesikhathi sokuhlola kanye nokuthola amasosha omzimba asezingeni eliphezulu. Ingahlinzeka ngokuklama kwe-antigen, ukuhlanganiswa, nokuguqulwa, ukungatheleleki kwezilwane, ukuhlolwa kwe-B cell eyodwa ye-B cell, ukulandelana kweseli elilodwa.