Chii chinonzi Antibody Engineering?

Nerubatsiro rweinjiniya yekurwisa antibody, zvave zvichigoneka kushandura saizi yemamorekuru, pharmacokinetics, immunogenicity, kusunga hukama, kujeka uye kuita basa remasoja ekudzivirira chirwere. Mushure mekugadzira masoja ekudzivirira chirwere, kusungirirwa kwemasoja ekudzivirira chirwere kunoita kuti ave akakosha zvakanyanya mukuongororwa kwekiriniki uye kurapwa. Kuburikidza neantibody engineering, vanogona kusangana nezvinodiwa zvemishonga uye yekuongorora budiriro yekutanga.

Chinangwa cheinjiniya yeantibody kugadzira uye kugadzira mabasa akanyanya, akagadzikana ayo masoja ekudzivirira chirwere asingakwanise kuwana, achiisa hwaro hwekugadzirwa kwemasoja ekudzivirira chirwere.

Alpha Lifetech, ine ruzivo rwayo rwakakura rweprojekiti muinjiniya yeantibody, inogona kupa yakagadziridzwa monoclonal uye polyclonal antibody masevhisi emarudzi akawanda, pamwe ne phage inoratidza antibody raibhurari kuvaka uye kuongorora masevhisi. Alpha Lifetech inogona kupa vatengi mhando yebiosimilar antibodies uye recombinant mapuroteni zvigadzirwa, pamwe nemasevhisi anowirirana, kugadzira masoja ekudzivirira chirwere anoshanda, akajeka, uye akagadzikana. Nekushandisa yakazara antibody, mapuratifomu eprotein uye phage kuratidza masisitimu, isu tinopa masevhisi anofukidza kumusoro nekudzika kwekugadzirwa kweanorwisa utachiona, anosanganisira masevhisi ehunyanzvi akadai seantibody humanization, kucheneswa kweantibody, kutevedzana kwevanhu, uye kusimbiswa kweantibody.

Nhanho yekupayona yeinjiniya yeantibody ine hukama nematekinoroji maviri:

--Recombinant DNA tekinoroji

--Hybridoma tekinoroji

Iko kukurumidza kusimudzira kweantibody engineering inoenderana neakakosha matatu matekinoroji:

-Gene cloning tekinoroji uye polymerase chain reaction

--Protein kutaura: Recombinant mapuroteni anogadzirwa nekutaura masisitimu senge mbiriso, mavhairasi akafanana netsvimbo, uye zvirimwa.

--Computer yakabatsira dhizaini dhizaini

Chizvarwa chekutanga chemasoja ekudzivirira chirwere chakaitwa vanhu kuti chigadzirwe chimeric antibodies, uko nharaunda inoshanduka yemonoclonal antibodies yakabatanidzwa kune inogara dunhu revanhu IgG mamorekuru. Iyo antigen inosunga dunhu (CDR) yechizvarwa chechipiri mbeva monoclonal antibody yakasimwa muIgG yemunhu. Kunze kwedunhu reCDR, mamwe masoja ekudzivirira chirwere anenge masoja emunhu, uye kuedza kwakaitwa kudzivirira kukonzeresa mhinduro dzehuman anti mouse antibody (HAMA) kana pachishandiswa masoja ekudzivirira chirwere chembeva pakurapwa kwevanhu.

 

Kuvaka raibhurari yekuratidzira phage, danho rekutanga nderekutora majini encoding antibodies, anogona kuparadzaniswa kubva kuB masero emhuka dzakabaiwa (immune raibhurari kuvaka), inotorwa zvakananga kubva kumhuka dzisina kubaiwa (yakasikwa raibhurari kuvaka), kana kunyange yakaunganidzwa mu vitro ine antibody gene zvidimbu (synthetic raibhurari kuvaka). Zvadaro, majini anowedzerwa nePCR, anopinzwa mumaplasmids, uye anoratidzirwa mugadziriro dzakakodzera dzevaenzi (yeast expression (kazhinji Pichia pastoris), prokaryotic expression (kazhinji E. coli), mammalian cell expression, plant cell expression, uye kutaura kwesero rezvipembenene zvine mavhairasi akafanana netsvimbo). Inonyanya kuzivikanwa ndeyeE. coli expression system, iyo inosanganisa encoding antibody sequence pafeji uye inokodha imwe yemapuroteni ephage (pIII kana pVIII). Iyo gene fusion ye, Uye inoratidzwa pamusoro peiyo bacteriophages. Iyo yakakosha yeiyi tekinoroji kugadzira raibhurari yekuratidzira phage, iyo ine mukana pamusoro pemaraibhurari echisikigo mukuti inogona kuve neinosunga. Mushure mezvo, masoja ekudzivirira chirwere ane antigen chaiwo anoongororwa kuburikidza nemaitiro ekusarudzwa kwehupenyu, maantigen anonangwa anogadziriswa, maphage asina kusungwa anogezwa kakawanda, uye phage dzakasungwa dzinogezwa kuti dziwedzere kupfumiswa. Mushure mekutenderera katatu kana kupfuura kwekudzokorora, hunyanzvi hwepamusoro uye masoja ekudzivirira epamusoro anoparadzaniswa.

Fig 3: Antibody Library Kuvaka uye Kuongorora

Recombinant DNA tekinoroji inogona kushandiswa kugadzira zvidimbu zveantibody. Masoja ekudzivirira chirwere anogona kutanga aitwa hydrolyzed negastric protease kugadzira (Fab ') zvidimbu zviviri, zvinozogayiwa nepapain kugadzira zvidimbu zveFab. Fv fragment ine VH neVL, iyo isina kugadzikana kwakashata nekuda kwekushaikwa kwema bond e disulfide. Nokudaro, VH neVL zvakabatanidzwa pamwe chete kuburikidza nepeptide pfupi ye15-20 amino acids kuti igadzire cheni imwe chete yakasiyana-siyana fragment (scFv) antibody ine molecular huremu hunosvika 25KDa.

Ongororo yemaitiro emasoja ekudzivirira chirwere muCamelidae (Camel, LIama, neAlpaca) yakaburitsa kuti masoja ekudzivirira chirwere anongori nemacheni anorema uye asina cheni dzakareruka, nekudaro anodaidzwa kunzi heavy chain antibodies (hcAb). The variable domain of heavy chain antibodies inonzi single domain antibodies kana nanobodies kana VHH, ine saizi ye12-15 kDa. Semamonomers, haana disulfide zvisungo uye akadzikama kwazvo, ane hukama hwakanyanya hwemaantigen.

Fig 5: Heavy Chain Antibody uye VHH/ Nanobody

Sero rakasununguka kutaura rinoshandisa matauriro eDNA yakasikwa kana kuti yakagadzirwa kuti iwane in vitro protein synthesis, kazhinji uchishandisa E. coli expression system. Iyo inogadzira mapuroteni nekukurumidza uye inodzivirira metabolic uye cytotoxic mutoro pamasero kana ichigadzira yakakura mapuroteni anobatana mu vivo. Inogonawo kugadzira mapuroteni akaoma kugadzirwa, seaya anonetsa kushandura mushure mekushandura kana kugadzira membrane mapuroteni.