Pulatifomu Yopangira Ma Antibody Awiriawiri

Alpha Lifetech ikhoza kupatsa makasitomala njira yodziwira bwino za kupezeka kwa ma antibodies ndi zinthu ndi ntchito za mapuloteni ophatikizana. Tikhoza kukonza ma antibodies ndi mphamvu zambiri, kudziwika bwino, komanso kukhazikika bwino. Alpha Lifetech ili ndi zida zosiyanasiyana zoyeretsera ma antibodies, zomwe zingapereke ntchito zoyeretsera ma antibodies kuchokera kuzinthu zosiyanasiyana monga ma antibodies a monoclonal a kalulu, nkhosa, nkhuku, ndi mbewa, komanso ntchito zoyeretsera ma protein A/G affinity ndi ntchito zoyeretsera ma antibodies. Kutengera ndi kapangidwe ka pulatifomu yonse ya pulatifomu yopezera ma antibodies, puloteni, ndi zina zotero, timaphimba ntchito zopanga ma antibodies pamwamba ndi pansi, ndipo timapereka ntchito zaukadaulo kuchokera ku kukonzekera ma antibodies, kuyeretsa ma antibodies a bispecific ndi kulekanitsa ma antibodies, kutsata ma antibodies, kutsimikizira ma antibodies, ndi zina zotero, zomwe zingagwiritsidwe ntchito pochiza ma antibodies a bispecific.

Ma antibodies a monoclonal amatanthauza ma immunoglobulins opangidwa ndi selo limodzi la B lomwe lili ndi ma antigen ambiri kapena ma epitopes. Kupanga ma antibodies a monoclonal poyamba kunakonzedwa pogwiritsa ntchito ukadaulo wa hybridoma kuti apange ma antibodies a mbewa. Makamaka, pophatikiza maselo a ndulu kuchokera ku mbewa zomwe zapatsidwa katemera ndi maselo a myeloma kuchokera kwa anthu kapena mbewa, maselo a hybridoma amapangidwa, omwe amatulutsa ma antibodies enaake. Ma antibodies a monoclonal opangidwa ndi mbewa amagwiritsidwa ntchito kwambiri pakufufuza nyama ndi kuzindikira matenda pambuyo pa ntchito zoyeretsera. Komabe, thupi la munthu limapanga yankho la chitetezo chamthupi ku mapuloteni a mbewa zakunja, kotero pali zoletsa zazikulu pakugwiritsira ntchito kwa ma antibodies a monoclonal a mbewa. Pambuyo pake, anthu adagwiritsa ntchito "humanization" kukonzekera ma antibodies a monoclonal kuchokera ku mbewa, ndipo adagwiritsa ntchito ukadaulo wa majini kuti asinthe ma antibodies a mbewa kuti akhale ndi gawo lokhazikika la immunoglobulin ya anthu kuti achepetse mphamvu yawo yoteteza thupi. Mtundu uwu wa antibody umadziwikanso kuti humanized monoclonal antibody. Kuphatikiza apo, kugwiritsa ntchito maselo a anthu kupanga ma antibodies a monoclonal kumatchedwa kukonzekera kwathunthu kwa ma antibodies a anthu a monoclonal. Ma antibodies a monoclonal amagwiritsidwa ntchito kwambiri mu ntchito zamankhwala ndi zamankhwala. Mu kafukufuku wazachipatala, ma antibodies a monoclonal angagwiritsidwe ntchito ngati ma immunomodulators, ma antibodies a monoclonal othandizira angagwiritsidwe ntchito pa mankhwala a khansa ndi matenda opatsirana ndi mavairasi, ndipo kupanga ma antibodies a monoclonal omwe amagwiritsidwa ntchito makamaka pochiza khansa.

Chiyambi cha Bispecific Antibody

Mu 1960, lingaliro la ma antibodies a bispecific linaperekedwa. Ma antibodies a bispecific, omwe amadziwikanso kuti ma antibodies a bispecific monoclonal, ndi ma antibodies opangidwa mwaluso pogwiritsa ntchito ukadaulo wa majini. Monga ma antibodies opangidwa mwaluso, ma antibodies a bispecific nthawi zambiri amakhala m'gulu la IgG ndipo amakhala ndi chidutswa chomangira ma antigen chomwe chimayang'ana gawo la CD3. Ma antibodies a bispecific ali ndi malo awiri enieni omangira ma antigen omwe amatha kumangirira ndikuzindikira ma antigen awiri osiyana nthawi imodzi, kapena ma epitopes awiri osiyana a antigen. Poyerekeza ndi ma antibodies a monoclonal, ma antibodies a bispecific ali ndi malo ena apadera omangira ma antigen, motero amakhala ndi luso lamphamvu komanso luso lolunjika, lomwe lingathe kulunjika molondola maselo a chotupa ndikuchepetsa poizoni womwe sunaganizidwe. Ma antibodies a bispecific amatha kugwira ntchito zosiyanasiyana zamoyo nthawi imodzi, monga kusonkhanitsa maselo a chitetezo chamthupi, kutseka njira zolumikizirana, ndikupha mwachindunji maselo a chotupa. Ma antibodies oyambirira a bispecific anakonzedwa makamaka kudzera mu kuphatikiza kwa mankhwala kapena kusakanikirana kwa maselo, koma njira iyi mwina idapita patsogolo pang'onopang'ono chifukwa cha kuphatikiza kosasinthika komanso kuvutika kupatula kuphatikiza komwe kumayang'aniridwa. Ndi chitukuko chopitilira cha ukadaulo wa majini, nsanja zambiri zatsopano zaukadaulo zapangidwa, monga ma knots in holes (KIH), CrossMab, DVD Ig, ndi zina zotero. Mapulatifomu awa amathetsa mavuto monga kusalingana kwa unyolo wolemera ndi unyolo wopepuka, ndikuwonjezera kufanana ndi kupangidwa kwa ma antibodies a bispecific.

Chithunzi 1 Chidule chosavuta cha njira zomwe zaperekedwa zogwirira ntchito ma antibodies a bispecific (bsAbs) mu mayeso azachipatala a oncology.(Chithunzi chochokera: Kuwunikanso kwa ma antibodies a bispecific ndi ma antibodies mu oncology ndi zovuta zachipatala - ScienceDirect)

Ukadaulo Wopanga Ma Antibodies Awiri

Njira zazikulu zopangira ma antibody awiriawiri ndi monga kuphatikiza mankhwala, ma source hybridoma anayi, ndi kukonzekera ma genetic engineering antibody. Pakati pawo, njira yolumikizira mankhwala imaphatikiza zidutswa ziwiri za ma antibody awiri a IgG kapena awiri a F (ab ') 2 kukhala ma antibodies awiriawiri pogwiritsa ntchito ma chemical coupling agents monga phthalimide ndi dithioacylbenzoic acid. Njirayi ndi yosavuta komanso yosavuta kugwiritsa ntchito, koma ingawononge malo omangira ma antigen, kuchepetsa ntchito ya ma antibodies, ndipo coupling agent yokha ilinso ndi kuchuluka kwa khansa. Njira zinayi za ma source hybridoma zimachokera pa kusakanikirana kwa maselo a somatic kuchokera ku mizere iwiri yosiyana ya ma hybridoma cell kuti iwonetse IgG ya mbewa yofananira. Kudzera mu ukadaulo wa majini, ma antibodies amatha kusinthidwa majini kuti apange ma antibodies awiriawiri. Ma antibodies awiri osiyana a monoclonal adapangidwa, ndipo zidutswa za Fab kapena zigawo zosinthika za unyolo wolemera ndi unyolo wopepuka wa ma antibodies awiriwa zidadulidwa padera. Kudzera mu cross-linking reaction kapena chain recombination technology, zidutswa ziwirizi zidaphatikizidwa kuti zipange bispecific antibody. Ngakhale kuti ukadaulo wa majini ndi wovuta kwambiri, pakadali pano ndiyo njira yomwe imagwiritsidwa ntchito kwambiri popanga ma antibody awiriawiri kuti asinthe kapangidwe ndi ntchito ya ma antibodies. Popanga ma antibody awiriawiri, mfundo ya ma antibody cross reactivity ingagwiritsidwe ntchito, koma ma antibody cross reactivity angayambitse zochitika zosakhudzana ndi zenizeni, kotero idzaganiziridwa mosamala pakugwiritsa ntchito mapangidwe a ma antibody awiriawiri, monga chithandizo cha ma antibody awiriawiri.

Kuyeretsa Ma Antibodies Awiriawiri

Kuyeretsa ma antibodies a Bispecific ndi njira yolekanitsira ndi kuyeretsa ma antibodies a high-purity target. Njira ziwiri, centrifugation ndi deep filtration, zimagwiritsidwa ntchito kuchotsa zonyansa zina zosungunuka. Antibody ya bispecific target imagwidwa poyamba ndi affinity chromatography. Kwa ma antibodies a IgG monga bispecific antibodies, protein A affinity chromatography imagwiritsidwa ntchito, pomwe kwa ma antibodies a non IgG monga bispecific antibodies, light chain based affinity chromatography ingagwiritsidwe ntchito. Pambuyo pake, antibody imayikidwa pansi pa pH yotsika kwa nthawi inayake, zomwe zimapangitsa kuti kapangidwe ka mapuloteni pamwamba pa envelopu ya kachilomboka kasokonezeke, motero kutaya mphamvu yopatsira maselo. Kusefa kwakuya kwapakati kumachitika kuti kuchotsenso zonyansa monga mapuloteni a cell host (HCP). Kuyera kwa ma antibodies kumawongoleredwa kudzera munjira monga ion exchange chromatography, ndipo ma virus aliwonse otsala amachotsedwa kudzera mu nanofiltration kapena ultrafiltration techniques. Pomaliza, chitsanzocho chimayikidwa ndikusinthidwa ndi formula buffer yoyenera.

Kayendedwe ka Ntchito Yopangira Ma Antibodies Awiriawiri

Masitepe	Zamkati mwa Utumiki	Mndandanda wa Nthawi
Kupanga Majini	Kupanga njira zitatu zosiyana zopangira ma sequences, kupanga ma plasmids a DNA antibody opangidwanso	Masabata awiri kapena atatu
Mayeso Ang'onoang'ono	Kupanga mapuloteni a bispecific antibody → Kuchuluka kochepa kwa maselo a zoyamwitsa → Kutsimikizika ndi SDS-PAGE → Kusanthula kolumikizana ndi ma antigen a mapuloteni ogwirizana ndi ELISA → Kuyerekeza kwa ma clones ndi ma antibodies a makolo	Masabata 5-6
Dziwani	Kufotokozera kwa ma antibodies awiri oyamba a bispecific → Kusanthula kogwirizana ndi ELISA → Kuwunika kwa ELISA kwa ma antibodies omwe amadziwika ndi ma antibodies a anti mbewa	Sabata imodzi
Kupanga Ma Antibody	Kupanga ma antibodies ambiri	Masabata 3-4

Ubwino Wathu

Timapereka chithandizo chapamwamba ndi chapansi pa madzi chokhudza kupanga ma antibodies, ndipo timapereka chithandizo chaukadaulo kuyambira kukonzekera ma antibodies, kuyeretsa ma antibodies a bispecific ndi kulekanitsa ndi kuyeretsa ma antibodies, kutsata ma antibodies, kutsimikizira ma antibodies, ndi zina zotero, zomwe zingagwiritsidwe ntchito pochiza ma antibodies a bispecific.

Kuwonetsa Kwambiri

Kuchuluka kwa mapuloteni m'maselo a CHO.
Malo Osungirako Okhazikika

> Masabata awiri mu seramu pa 37 °C.
Palibe Vuto Losungunula

>30 mg/ml
Maselo Okhazikika

Ma cell ambiri okhazikika okhala ndi ma antigen a chotupa apangidwa kuti atsimikizire ma antibodies ambiri a bispecific