Kodi Uinjiniya wa Antibody ndi Chiyani?

Mothandizidwa ndi mainjiniya a ma antibodies, zatheka kusintha kukula kwa mamolekyulu, pharmacokinetics, immunogenicity, binding affinity, specificity ndi effector function ya ma antibodies. Pambuyo popanga ma antibodies, kumangirira kwa ma antibodies kumawapangitsa kukhala ofunika kwambiri pa matenda ndi chithandizo chamankhwala. Kudzera mu mainjiniya a ma antibodies, amatha kukwaniritsa zosowa za mankhwala ndi matenda oyambirira.

Cholinga cha kupanga ma antibodies ndikupanga ntchito zenizeni komanso zokhazikika zomwe ma antibodies achilengedwe sangathe kuchita, ndikuyika maziko opanga ma antibodies ochiritsira.

Alpha Lifetech, yokhala ndi chidziwitso chachikulu pa ntchito yake yokonza ma antibody, ingapereke ntchito zodzitetezera za monoclonal ndi polyclonal kwa mitundu yosiyanasiyana ya ma antibodies, komanso ntchito zomanga ndi kuyesa laibulale ya ma antibody a phage display. Alpha Lifetech ikhoza kupatsa makasitomala ma antibodies abwino a biosimilar ndi zinthu zopangidwa ndi mapuloteni ogwirizana, komanso ntchito zofanana, kuti apange ma antibodies ogwira mtima, apadera kwambiri, komanso okhazikika. Pogwiritsa ntchito ma antibody ambiri, ma protein platforms ndi ma phage display system, timapereka ntchito zokhudzana ndi kupanga ma antibody pamwamba ndi pansi, kuphatikizapo ntchito zaukadaulo monga kupanga ma antibody kukhala munthu, kuyeretsa ma antibody, kutsata ma antibody, ndi kutsimikizira ma antibody.

Gawo loyamba la uinjiniya wa ma antibody likugwirizana ndi matekinoloje awiri:

--Ukadaulo wa DNA wophatikizana

--Ukadaulo wa Hybridoma

Kukula mwachangu kwa ma antibodies kumakhudzana ndi ukadaulo wofunikira atatu:

--Ukadaulo wa cloning wa majini ndi polymerase chain reaction

--Kuchuluka kwa mapuloteni: Mapuloteni ophatikizana amapangidwa ndi machitidwe owonetsera monga yisiti, mavairasi ooneka ngati ndodo, ndi zomera

--Kapangidwe ka kapangidwe ka makompyuta kothandizidwa ndi makompyuta

Mbadwo woyamba wa ma antibodies unasinthidwa kukhala waumunthu kuti upange ma antibodies a chimeric, komwe dera losinthika la ma antibodies a monoclonal a mbewa linalumikizidwa ndi dera lokhazikika la mamolekyu a IgG a anthu. Dera lomangira ma antigen (CDR) la antibody ya monoclonal ya mbewa ya m'badwo wachiwiri linaikidwa mu IgG ya anthu. Kupatula dera la CDR, ma antibodies ena onse ndi pafupifupi ma antibodies a anthu, ndipo khama linapangidwa kuti apewe kuyambitsa mayankho a antibody ya mbewa ya anthu (HAMA) pogwiritsa ntchito ma antibodies a mbewa pochiza anthu.

 

Kuti mupange laibulale yowonetsera phage, gawo loyamba ndikupeza majini omwe amalemba ma antibodies, omwe amatha kuchotsedwa ku maselo a B a nyama zomwe zapatsidwa katemera (kumanga laibulale ya chitetezo chamthupi), kuchotsedwa mwachindunji kuchokera ku nyama zomwe sizinalandire katemera (kumanga laibulale yachilengedwe), kapena kusonkhana mu vitro ndi zidutswa za majini a antibody (kumanga laibulale yopangidwa). Kenako, majiniwo amakulitsidwa ndi PCR, kuyikidwa mu plasmids, ndikufotokozedwa mu machitidwe oyenera a host (kutulutsa yisiti (nthawi zambiri Pichia pastoris), kuonetsa prokaryotic (nthawi zambiri E. coli), kuonetsa maselo a mammalia, kuonetsa maselo a zomera, ndi kuonetsa maselo a tizilombo omwe ali ndi kachilombo ka ndodo). Chodziwika kwambiri ndi dongosolo lowonetsa E. coli, lomwe limaphatikiza njira yeniyeni yolembera ma antibodies pa phage ndikulemba imodzi mwa mapuloteni a phage shell (pIII kapena pVIII). Kuphatikizika kwa majini, Ndipo kumawonetsedwa pamwamba pa bacteriophages. Pakati pa ukadaulo uwu ndikupanga laibulale yowonetsera phage, yomwe ili ndi ubwino kuposa malaibulale achilengedwe chifukwa imatha kukhala ndi zomangira zinazake. Pambuyo pake, ma antibodies omwe ali ndi antigen specificity amafufuzidwa kudzera mu njira yosankha yachilengedwe, ma antigen omwe akufuna amakhazikika, ma phages osamangidwa amatsukidwa mobwerezabwereza, ndipo ma phages omangidwa amatsukidwa kuti awonjezere mphamvu. Pambuyo pa maulendo atatu kapena kuposerapo, ma antibodies omwe ali ndi specificity yapamwamba komanso high affinity amachotsedwa.

Chithunzi 3: Kumanga ndi Kuwunika Laibulale ya Antibody

Ukadaulo wa DNA wophatikizana ungagwiritsidwe ntchito kupanga zidutswa za antibody. Ma antibodies a Fab poyamba amatha kusungunuka ndi gastric protease kuti apange zidutswa ziwiri (Fab '), zomwe kenako zimagayidwa ndi papain kuti apange zidutswa za Fab payokha. Chidutswa cha Fv chimakhala ndi VH ndi VL, zomwe zimakhala ndi kukhazikika kosakhazikika chifukwa chosowa ma disulfide bond. Chifukwa chake, VH ndi VL zimalumikizidwa pamodzi kudzera mu peptide yayifupi ya 15-20 amino acid kuti apange antibody imodzi yosinthika ya unyolo (scFv) yokhala ndi kulemera kwa molekyulu pafupifupi 25KDa.

Kafukufuku wa kapangidwe ka ma antibodies mu Camelidae (Camel, LIama, ndi Alpaca) wasonyeza kuti ma antibodies ali ndi ma chain olemera okha ndipo alibe ma chain opepuka, motero amatchedwa ma heavy chain antibodies (hcAb). Malo osinthika a ma heavy chain antibodies amatchedwa single domain antibodies kapena nanobodies kapena VHH, okhala ndi kukula kwa 12-15 kDa. Monga ma monomers, alibe ma disulfide bonds ndipo ndi okhazikika kwambiri, okhala ndi mphamvu yayikulu kwambiri ya ma antigen.

Chithunzi 5: Woteteza Unyolo Wolemera ndi VHH/ Nanobody

Kusalankhulana kwa maselo kumagwiritsa ntchito kufotokozedwa kwa DNA yachilengedwe kapena yopangidwa kuti ipange mapuloteni a in vitro, nthawi zambiri pogwiritsa ntchito njira yofotokozera ya E. coli. Imapanga mapuloteni mwachangu ndipo imapewa kupsinjika kwa kagayidwe kachakudya ndi poizoni m'maselo popanga mapuloteni ambiri obwerezabwereza m'thupi. Imathanso kupanga mapuloteni omwe ndi ovuta kupanga, monga omwe ndi ovuta kusintha pambuyo powasintha kapena kupanga mapuloteni a nembanemba.