Chii chinonzi Antibody Engineering?

Nerubatsiro rwekugadzira maantibody, zvave zvichikwanisika kugadzirisa saizi yemamorekuru, pharmacokinetics, immunogenicity, binding affinity, specificity uye effector function yemaantibody. Mushure mekugadzira maantibody, kubatana kwacho kweantibody kunoita kuti ave akakosha zvikuru mukuongorora nekurapa. Kuburikidza nekugadzira maantibody, anogona kusangana nezvinodiwa zvekutanga kwemishonga nekuongorora.

Chinangwa chekugadzira maantibody ndechekugadzira nekugadzira mabasa akasimba uye akanyatsojeka ayo maantibody echisikigo asingagoni kuitwa, zvichiisa hwaro hwekugadzirwa kwemaantibody ekurapa.

Alpha Lifetech, ine ruzivo rwakakura mukugadzira maantibody, inogona kupa masevhisi ekudzivirira maantibody emhando dzakasiyana-siyana, pamwe nemabasa ekuvaka nekuongorora maantibody emhando yepamusoro ekugadzira maantibody emhando yepamusoro ekugadzira maantibody emhando yepamusoro. Alpha Lifetech inogona kupa vatengi maantibody emhando yepamusoro uye zvigadzirwa zveprotein recombinant, pamwe nemasevhisi anoenderana, kuti vagadzire maantibody anoshanda zvakanaka, akanyatsotsanangurwa, uye akasimba. Nekushandisa maantibody akakwana, mapuratifomu eprotein uye masisitimu ekuratidza maantibody emhando yepamusoro, tinopa masevhisi anofukidza kugadzirwa kwemaantibody emhando yepamusoro nepasi, kusanganisira masevhisi ehunyanzvi akadai sekuita kuti maantibody ave munhu, kucheneswa kwemaantibody, kutevedzana kwemaantibody, uye kusimbiswa kwemaantibody.

Danho rekutanga rekugadzira maantibody rine chekuita nematekinoroji maviri:

--Tekinoroji yeDNA inosanganiswa

--Tekinoroji yeHybridoma

Kukurumidza kukura kwekugadzirwa kwemaantibody kwakabatana nematekinoroji matatu akakosha:

--Tekinoroji yekuumba majini uye reaction ye polymerase chain

--Kuonekwa kweprotein: Mapuroteni anowanikwazve anogadzirwa nemaitiro ekutaura akadai sembiriso, mavhairasi akaita setsvimbo, uye zvirimwa

--Dhizaini yekuvaka inobatsirwa nekombiyuta

Chizvarwa chekutanga chema antibodies chakagadzirwa semunhu kuti chigadzirwe ma antibodies e chimeric, uko nzvimbo inoshanduka yema antibodies e monoclonal embeva yakabatana nenzvimbo inogara iripo yema molecule e IgG emunhu. Nzvimbo inosunga maantigen (CDR) yechizvarwa chechipiri che mouse monoclonal antibody yakasimwa mu human IgG. Kunze kwenzvimbo ye CDR, mamwe ma antibodies ese anenge ari ma antibodies emunhu, uye kuedza kwakaitwa kudzivirira kukonzera mhinduro dze human anti mouse antibody (HAMA) pakushandisa ma antibodies e mouse clone pakurapa vanhu.

 

Kuti ugadzire raibhurari yekuratidza phage, danho rekutanga nderekuwana majini anochengetedza ma antibodies, anogona kuparadzaniswa kubva kumasero eB emhuka dzakadzivirirwa (kuvakwa kweraibhurari yekudzivirira muviri), anotorwa zvakananga kubva kumhuka dzisina kudzivirirwa (kuvakwa kweraibhurari yechisikigo), kana kutounganidzwa mu vitro nezvidimbu zvemajini ekudzivirira muviri (kuvakwa kweraibhurari yakagadzirwa). Zvadaro, majini anowedzerwa nePCR, anoiswa muplasmids, uye anoburitswa mumasystem akakodzera ekugamuchira (yeast expression (kazhinji Pichia pastoris), prokaryotic expression (kazhinji E. coli), mammalia cell expression, plant cell expression, uye insect cell expression ine utachiona hwemavhairasi akaita setsvimbo). Yakajairika ndiyo E. coli expression system, iyo inobatanidza specific encoding antibody sequence paphage uye inochengetedza imwe yemapuroteni ephage shell (pIII kana pVIII). Kusanganiswa kwemajini, Uye kunoratidzwa pamusoro pemabacteriophages. Chinhu chikuru chetekinoroji iyi kuvaka raibhurari yekuratidza phage, ine mukana pamusoro pemaraibhurari echisikigo nekuti inogona kuva nekusunga kwakananga. Zvadaro, ma antibodies ane antigen specificity anoongororwa kuburikidza nemaitiro ekusarudza emuviri, ma target antigen anogadziriswa, ma unbound phages anogezwa kakawanda, uye ma bound phages anogezwa kuti awedzere kuwedzerwa. Mushure mekudzokorora katatu kana kupfuura, ma high specificity uye high affinity antibodies anobviswa.

Mufananidzo 3: Kuvaka uye Kuongorora Raibhurari yeAntibody

Tekinoroji yeDNA recombinant inogona kushandiswa kugadzira zvidimbu zveantibody. Ma antibodies eFab anogona kutanga angonyungudutswa negastric protease kuti agadzire zvidimbu zviviri (Fab '), izvo zvinozogayiwa nepapain kuti agadzire zvidimbu zveFab zvega zvega. Chidimbu cheFv chine VH neVL, izvo zvisina kugadzikana zvakanaka nekuda kwekushaikwa kwe disulfide bonds. Nokudaro, VH neVL zvakabatana pamwe chete kuburikidza ne peptide pfupi ye 15-20 amino acids kuti vagadzire single chain variable fragment (scFv) antibody ine huremu hwemamorekuru hunosvika 25KDa.

Chidzidzo chechimiro cheantibody muCamelidae (Camel, LIama, uye Alpaca) chakaratidza kuti ma antibodies ane cheni dzinorema chete uye haana cheni dzakareruka, saka anonzi ma heavy chain antibodies (hcAb). Nzvimbo inoshanduka yema heavy chain antibodies inonzi single domain antibodies kana nanobodies kana VHH, ine saizi ye12-15 kDa. Sema monomers, haana disulfide bonds uye akagadzikana zvikuru, ane hukama hwakanyanya hwema antigen.

Mufananidzo 5: Heavy Chain Antibody uye VHH/ Nanobody

Kusununguka kwemasero kunoshandisa kuburitswa kweDNA yechisikigo kana yekugadzira kuti iwane kugadzirwa kwemapuroteni mu vitro, kazhinji uchishandisa sisitimu yekutaura yeE. coli. Inogadzira mapuroteni nekukurumidza uye inodzivirira mutoro we metabolic uye cytotoxic pamasero painogadzira mapuroteni akawanda e recombinant mumuviri. Inogonawo kugadzira mapuroteni akaoma kugadzira, akadai seaya akaoma kugadzirisa mushure mekushandurwa kana kugadzira mapuroteni e membrane.