Ipulatifomu Yokuthuthukiswa Kwama-Antibody E-Phage Display

I-Alpha Lifetech, enokuhlangenwe nakho kwayo okukhulu kwephrojekthi ekutholakaleni kwama-antibody, ingahlinzeka ngezinsizakalo zama-antibody e-monoclonal kanye ne-polyclonal ezenziwe ngokwezifiso zezinhlobo eziningi, kanye nezinsizakalo zokwakha kanye nokuhlola umtapo wezincwadi wama-antibody. I-Alpha Lifetech ingahlinzeka amakhasimende ngemikhiqizo yama-antibody kanye nama-antibody aphinde ahlanganiswe kabusha aqinisekisiwe ngekhwalithi, kanye nezinsizakalo ezifanayo, ukulungiselela ama-antibody asebenzayo, aqondile kakhulu, futhi azinzile. Ngokusebenzisa amapulatifomu ama-antibody aphelele kanye nobunjiniyela bama-antibody, sinikeza izinsizakalo ezihlanganisa ukukhiqizwa kwama-antibody okuphezulu nokwesibili, kufaka phakathi izinsizakalo zobuchwepheshe ezifana nokulungiselela ama-antibody, ukuhlanzwa, ukulandelana kwama-antibody, kanye nokuqinisekiswa.

I-Alpha Lifetech ineplatifomu yokuthola ama-antibody avuthiwe enikeza amakhasimende izinsizakalo ezahlukahlukene ezihlanganisa ukuzivikela kwezilwane, ukwakhiwa kanye nokuhlolwa komtapo wolwazi we-phage display antibody, ukulandelana kwama-antibody, ukuvezwa kwama-antibody, ukuhlanzwa kwama-antibody, ukuqinisekiswa kwama-antibody, kanye nokulebula ama-antibody ngokusekelwe kubuchwepheshe be-hybridoma, ubuchwepheshe be-single B cell, ubuchwepheshe bokubonisa ama-phage, nokuningi. I-Alpha Lifetech inomugqa wokukhiqiza ojwayelekile, futhi ngaphezu kwezinsizakalo zokulungiselela ama-antibody, izinsizakalo ezisekelayo ezifana nokwenza ama-antibody abe ngabantu, ukuvuthwa kwama-antibody affinity, ukwakheka kanye nokuthuthukiswa kwemithi ye-ADC, kanye nokwakhiwa kokulandelana okulandelayo kwe-CAR-T nakho kunganikezwa. Ngesikhathi esifanayo, i-Alpha Lifetech yakhe umtapo wolwazi we-antibody ngokusekelwe kumapulatifomu obuchwepheshe bokubonisa ama-phage e-M13, T4, kanye ne-T7, enomthamo wokugcina ofinyelela ku-10^8-10^9. Izinga elihle, izinga lokufakwa, kanye nokuhlukahluka komtapo wolwazi konke kungafinyelela ngaphezu kuka-90%.

Ubuchwepheshe bokubonisa i-Phage

Ukukhiqizwa kwama-antibodies e-monoclonal kwaqale kwalungiselelwa kusetshenziswa ubuchwepheshe be-hybridoma monoclonal antibody ukukhiqiza i-monoclonal antibody yegundane i-hybridoma. Ngokukhethekile, ngokuhlanganisa amaseli e-spleen avela kumagundane agonyiwe namaseli e-myeloma avela kubantu noma emagundwini, kwakheka amaseli e-hybridoma, akhipha ama-antibodies athile e-hybridoma monoclonal antibody. Ngokuguqulwa kwama-antibodies egundane e-monoclonal okwenziwe ngabantu kanye nokuguqulwa kobunjiniyela bezakhi zofuzo zama-antibodies ukuze anikezwe indawo eqhubekayo ye-immunoglobulin yomuntu, ukuqina kwawo kwe-immunogenicity kungancishiswa. Ubunjiniyela be-monoclonal antibody busetshenziswa kabanzi ekusetshenzisweni kwezokwelapha kanye nokwelashwa.

I-Alpha Lifetech ingahlinzeka ngezinsizakalo zobunjiniyela be-antibody eyodwa kusukela ekwakhiweni kwephrothokholi yocwaningo lwangaphambi kokwelashwa kuya ekutholakaleni kwama-conjugates ezidakamizwa zama-antibody (ama-ADC) kanye nokuqinisekiswa kwezilwane. Ama-conjugates ezidakamizwa zama-antibody (ama-ADC) angaletha imithi ye-chemotherapy kumaseli omdlavuza. Ngemva kokubopha ezinhlosweni ezithile ezivezwe kumaseli omdlavuza, i-ADC ikhipha imithi ye-cytotoxic kumaseli omdlavuza. I-Alpha Lifetech ingahlinzeka namakhasimende ngezinsizakalo eziphelele zokuvuthwa kwe-antibody affinity. Ngokuthuthukiswa kokuguqulwa kwe-mutation kanye nobuchwepheshe bokuhlola i-phage ephezulu, ama-antibodies ane-affinity ethile ahlolwa kuqala, bese kuthi ama-mutations e-amino acid aqalwe ukukhiqiza izinhlobo ezahlukene. Ngemva kwalokho, ama-antibodies ane-affinity ephezulu ahlolwa futhi ahlolwa kusetshenziswa amasu okuhlola. Ngemva kwemijikelezo eminingi yokwenza ngcono kanye nokuhlaziywa kwesakhiwo, ama-antibodies ane-affinity ephezulu kanye nokucaciswa okuqinile ekugcineni atholakala.

I-Alpha Lifetech ingakha izinhlobo ezahlukene zemitapo yokubonisa i-antibody phage, okuhlanganisa imitapo yolwazi yokuzivikela komzimba, imitapo yolwazi yemvelo, imitapo yolwazi e-semi synthetic, kanye nemitapo yolwazi yokwenziwa. Ngokusekelwe kumthamo ophezulu wemitapo yolwazi yokuzivikela komzimba, ama-antibodies akhethekile kakhulu angatholakala. Ama-vector amaningi e-phagemid njenge-pMECS, i-pComb3X, kanye ne-pCANTAB 5E anganikezwa, kanye nezinhlobo ezifana ne-TG1 E. coli, i-XL1 Blue, kanye ne-ER2738. Ngaphezu komthamo womtapo wolwazi ofika ku-10^9, izinga lokufakwa kwezingcezu eziqondiwe zomtapo wolwazi nalo liphezulu, okudala izimo ezanele zokuhlola ama-antibodies anelise amakhasimende. Ishadi lokugeleza kwesevisi yokulungiselela ama-antibody ye-Alpha Lifetech esekelwe kubuchwepheshe bokubonisa ama-phage liboniswe kuMfanekiso 1.

Umfanekiso 1 Umdwebo wenqubo wokulungiselela ama-antibodies e-monoclonal asekelwe kubuchwepheshe bokubonisa i-phage.

Ukuhamba Komsebenzi Wokukhiqiza Ama-Antibody Esikrini Se-Phage Display

Izinyathelo	Okuqukethwe Kwesevisi	Umugqa wesikhathi
Isinyathelo 1: Ukugonywa kwezilwane	(1) Ukugonywa kwezilwane izikhathi ezine, ukugonywa kwe-booster umthamo owodwa, inani eliphelele lemithamo emi-5 egonyiwe. (2) I-serum engemihle ngaphambi kokugonywa yaqoqwa, futhi i-ELISA yenziwa kumthamo wesine ukuthola i-serum titer. (3) Uma i-serum antibody titer yomthamo wesine ihlangabezana nezidingo, umthamo owodwa owengeziwe wokugoma uzonikezwa ezinsukwini eziyi-7 ngaphambi kokuqoqwa kwegazi. Uma kungahlangabezani nezidingo, ukugoma okuvamile kuzoqhubeka. (4) Amandla afanelekile, ukuqoqwa kwegazi kanye nokuhlukaniswa kwama-monocyte	Amaviki ayi-10
Isinyathelo 2: Ukulungiselela i-cDNA	(1) I-PBMC Total RNA Extraction (Ikhithi Yokukhipha i-RNA) (2) Ukulungiswa kwe-RT-PCR okuthembekile kakhulu kwe-cDNA (ikhithi yokubhala engemuva)	Usuku olu-1
Isinyathelo 3: Ukwakhiwa Komtapo Wolwazi Wokulwa Namagciwane	(1) Kusetshenziswa i-cDNA njengethempulethi, izakhi zofuzo zandiswa ngemijikelezo emibili ye-PCR. (2) Ukwakhiwa nokuguqulwa kwe-Phage: i-gene splicing phagemid vector, ukuguqulwa kwe-electroporation kwamagciwane aphethe i-TG1, ukwakhiwa komtapo wezincwadi we-antibody. (3) Ukuhlonza: Khetha ngokungahleliwe ama-clone angu-24, izinga lokuhlonza i-PCR elihle + izinga lokufaka. (4) Ukulungiswa kwe-phage okusizwayo: Ukukhulisa i-phage ye-M13+ukuhlanzwa. (5) Ukuhlenga umtapo wolwazi we-Phage display	Amaviki angu-3-4
Isinyathelo 4: Ukuhlolwa Kwelabhulali Ye-Antibody (imizuliswano emi-3)	(1) Ukuhlolwa okuzenzakalelayo kwemizuliswano emithathu (ukuhlolwa kwesigaba esiqinile): ukuhlolwa kwengcindezi ukuze kususwe ama-antibodies angacacile ngezinga eliphezulu ngangokunokwenzeka. (2) Kukhethwe i-bacteriophage yokukhuliswa kwe-clone eyodwa+ukubonakaliswa okubangelwa yi-IPTG+ukutholwa kwe-ELISA kwama-clone amahle. (3) Wonke ama-clone amahle akhethiwe ukuze kuhlolwe izakhi zofuzo.	Amaviki angu-4-5

Inzuzo Yobuchwepheshe Bokubonisa I-phage

I-Alpha Lifetech inolwazi oluningi emkhakheni wokuthuthukiswa kwama-antibody. Eminyakeni edlule, i-Alpha Lifetech isungule ipulatifomu ephelele yokuthuthukiswa kwama-antibody.

Izinsizakalo Zokusekela

Singahlinzeka ngezinsizakalo ezahlukene zokwakha umtapo wolwazi wokuzivikela komzimba osekelwe ezilwaneni kanye nezinsizakalo zokuhlola umtapo wolwazi wokuzivikela komzimba wemvelo ngokuya ngezidingo zamakhasimende.

I-Target Eningi

Izinsizakalo eziningi zokuthola ama-antibody aqondiwe ziyatholakala: amaprotheni, ama-peptide, ama-molecule amancane, amagciwane, amaprotheni e-membrane, i-mRNA, njll.

Amavektha Amaningi

Insizakalo yokwakha umtapo wolwazi eyenzelwe wena, singahlinzeka ngamavekhtha ahlukahlukene e-bacteriophage kufaka phakathi i-PMECS, i-pComb3X, kanye ne-pCANTAB 5E, futhi siwashintshe ngokwezidingo zamakhasimende.

Ipulatifomu Yabadala

Umthamo wokugcina ungafinyelela ku-10 ^ 8-10 ^ 9, amazinga okufakwa angaphezulu kuka-90%, futhi ukuqina kwama-antibodies atholakala ngokuhlolwa ngokuvamile kusezingeni le-nM pM.

INSEVISI Ehlobene

Amasu Okuthuthukisa Ama-Antibody Amaningi

Isevisi Yokuthuthukiswa Kwama-Antibody E-Monoclonal

Ipulatifomu Yobuchwepheshe be-Hybridoma

Ipulatifomu Yokuhlunga Amaseli Amabili Amabili

Ipulatifomu Yokuthuthukiswa Kwama-Antibody E-Phage Display

ukuthuthukiswa kwe-antibody ye-monoclonal - i-Alpha Lifetech

Isevisi Yokuthuthukiswa Kwama-Antibody E-Monoclonal

Singahlinzeka ngezinsizakalo zokuthuthukiswa kwama-antibody e-monoclonal asezingeni eliphezulu, ahlanzekile kakhulu, futhi aqondile kakhulu, okuhlanganisa ukukhiqizwa kwama-antibody e-monoclonal egundane kanye nama-antibody e-rabbit monoclonal.

Ipulatifomu Yobuchwepheshe be-Hybridoma

Kubandakanya uhlelo lokugoma, izinsizakalo zokulungiselela ama-antibody, ukuhlanzwa kwama-antibody, ukulandelana kwe-antibody ephezulu, ukuqinisekiswa kwama-antibody, njll.

Ipulatifomu Yokuhlunga Amaseli Amabili Amabili

I-Alpha Lifetech inezinzuzo ngesikhathi sokuhlola kanye nokuthola ama-antibodies asezingeni eliphezulu. Ingahlinzeka ngomklamo we-antigen, ukwakheka, kanye nokuguqulwa, ukuzivikela kwezilwane, ukuhlolwa kokunothisa iseli elilodwa le-B, kanye nokulandelana kweseli elilodwa.