Basa reScFv reKuwana MaAntibody

Alpha Lifetech Inc.inodada nekupa mabasa akazara ekuvaka maraibhurari eScFv kuti isangane nezvinodiwa zvakasiyana-siyana zvesainzi yepasi rose. Chinangwa chedu ndechekunzwisisa nekuzadzisa zvinodiwa nevatengi vakasiyana uye kubatsira nematambudziko ari kuuya uye ari kubuda mubasa rekutsvagisa.

Chii chinonzi scFv- Single Chain Variable Fragment Antibody?

Antibody ine cheni imwe chete ine chimedu chechiedza chinoshanduka (VL) uye chimedu cheheavy (VH) chakabatanidzwa ne flexible peptide linker, chinogona kuratidzwa zviri nyore muchimiro chinoshanda pamusoro pe phage uye nekudaro chinowanzozivikanwa se single-chain variable fragment (scFv). Chimedu che scFv ndicho chikamu chidiki che immunoglobulin molecule (kunze kwema nanobodies) chine basa mumabasa ekubatanidza antigen uye chinobvumira mapuroteni ekugadzira kuvandudza hunhu hwe scFv (single chain fragment variable) zvakaita sekuwedzera kwehukama uye kushandurwa kwe specificity. Mu Alpha Lifetech Inc., ma antibodies e scFv akagadzirwa zvakanyanya kubva ku hybridoma, spleen cells kubva kumakonzo akadzivirirwa, uye B lymphocytes kubva kuvanhu. Total RNA kana mRNA inotanga yaparadzaniswa kubva ku hybridoma, spleen, lymph cells, uye bone marrow cells ichiteverwa ne reverse transcribed kuita cDNA kuti ishande se template ye antibody gene amplification (PCR).

Tekinoroji yeScFv Antibody Development-Phage Display

Kusiyana kweraibhurari kunoenderana nehuwandu hwema clones akazvimiririra. Kuti vagadzire raibhurari yakakura ye scFv antibody, masayendisiti edu achashandisa nzira yedu yematanho maviri yekugadzira raibhurari. Mudanho rekutanga remaitiro atsanangurwa, ma repertoire ekutanga anogadzirwa kubva kuzvigadzirwa zvePCR zvinonyora ma domain eVH, Vκ, uye Vλ, zvichiburitsa maraibhurari akajairika epakati (1 - 100 miriyoni ma clones). Mudanho rechipiri, zvidimbu zveVH (kana VL) zvinoparadzaniswa nekugaya plasmid DNA yakacheneswa kubva kuma repertoire ekutanga uye zvinounganidzwa mu acceptor phagemid vector ine ma repertoire eVL (kana VH). Iyi nzira itsva inowedzera hukuru hwemaraibhurari zvakanyanya (10).¹⁰-10¹²maclones).

Kuongororwa kweAntibody yeScFv yakakura uye ine simba rakawanda

Pakugadzirira scFv kwakakura, zviri pachena kuti kusarudzwa kwemugadziri wekutaura kuchave nemhedzisiro yakakura pakugadzirwa kwemaitiro, nzira dzekugadzirisa dzinotevera, uye mutengo wemaitiro. Kusarudzwa kwenzira yekugadzira kuchaenderana nekukosha kwekushandurwa kwemashure mekushandura uye nguva yekuvandudzwa kwekiriniki nehupfumi, izvo zvinonyanya kutsamirawo 'mugadziri wekutaura'. Hapana sisitimu yekutaura yepasi rose inovimbisa huwandu hwakawanda hwekutaura kwechigadzirwa kune akasiyana masoja ekudzivirira chirwere kana mapuroteni.