Kodi Antibody Engineering ndi chiyani?

Mothandizidwa ndi uinjiniya wa antibody, zakhala zotheka kusintha kukula kwa maselo, pharmacokinetics, immunogenicity, kumangiriza kuyanjana, kutsimikizika ndi magwiridwe antchito a ma antibodies. Pambuyo popanga ma antibodies, kumangiriza kwa ma antibodies kumawapangitsa kukhala ofunikira kwambiri pakuzindikira komanso kuchiza matenda. Kupyolera mu uinjiniya wa ma antibody, amatha kukwaniritsa zosowa zamankhwala komanso kuzindikira koyambirira.

Cholinga cha uinjiniya wa ma antibody ndikupanga ndi kupanga ntchito zenizeni, zokhazikika zomwe ma antibodies achilengedwe sangakwaniritse, ndikuyika maziko opangira ma antibodies achire.

Alpha Lifetech, ndi luso lake lalikulu la projekiti mu uinjiniya wa antibody, atha kupereka makonda amtundu wa monoclonal ndi polyclonal antibody pamitundu ingapo, komanso ntchito zowonetsera laibulale ya antibody ndi zowunikira. Alpha Lifetech ikhoza kupatsa makasitomala ma antibodies apamwamba a biosimilar ndi mapuloteni ophatikizananso, komanso ntchito zofananira, kuti apange ma antibodies abwino, odziwika bwino komanso okhazikika. Pogwiritsa ntchito ma antibody okwanira, mapulaneti a mapuloteni ndi makina owonetsera phage, timapereka chithandizo chokhudza kumtunda ndi kumunsi kwa antibody kupanga, kuphatikizapo ntchito zaukadaulo monga kulimbikitsa anthu, kuyeretsa ma antibody, kutsatizana kwa antibody, ndi kutsimikizira kwa antibody.

Gawo loyambitsa uinjiniya wa antibody likugwirizana ndi matekinoloje awiri:

--Recombinant DNA Technology

--ukadaulo wa Hybridoma

Kukula mwachangu kwa uinjiniya wa antibody kumagwirizana ndi matekinoloje atatu ofunikira:

--Gene cloning technology ndi polymerase chain reaction

--Manenedwe a mapuloteni: Mapuloteni ophatikizanso amapangidwa ndi machitidwe monga yisiti, ma virus owoneka ngati ndodo, ndi zomera.

--Kupanga kothandizidwa ndi makompyuta

M'badwo woyamba wa ma antibodies adapangidwa kuti apange ma antibodies a chimeric, pomwe gawo losinthika la ma antibodies a monoclonal adalumikizidwa ndi gawo lokhazikika la mamolekyu a IgG amunthu. Chigawo chomangirira cha antigen (CDR) cha m'badwo wachiwiri wa mbewa monoclonal antibody adasinthidwa kukhala IgG yamunthu. Kupatula dera la CDR, ma antibodies ena onse ndi pafupifupi ma antibodies aumunthu, ndipo zoyesayesa zidapangidwa kuti tipewe kuyankha mayankho a anti mouse antibody (HAMA) pogwiritsa ntchito ma antibodies a mbewa pochiza anthu.

 

Kuti mupange laibulale yowonetsera phage, gawo loyamba ndikupeza ma jini ophatikizira ma antibodies, omwe amatha kupatulidwa kuchokera ku maselo a B a nyama zotetezedwa (kumanga laibulale ya chitetezo chamthupi), yotengedwa mwachindunji kuchokera ku nyama zopanda katemera (zomangamanga zama library), kapenanso zosonkhanitsidwa mu vitro ndi zidutswa za jini za antibody (kupanga laibulale yopangira). Kenaka, majini amakulitsidwa ndi PCR, amalowetsedwa mu plasmids, ndipo amasonyezedwa m'machitidwe oyenerera osungira (yeast expression (kawirikawiri Pichia pastoris), mawu a prokaryotic (kawirikawiri E. coli), mawonekedwe a maselo a mammalian, mawonekedwe a maselo a zomera, ndi mawonekedwe a selo la tizilombo omwe ali ndi mavairasi opangidwa ndi ndodo). Chofala kwambiri ndi E. coli expression system, yomwe imaphatikiza ma encoding antibody motsatizana pa phage ndikuyika imodzi mwa mapuloteni a chipolopolo cha phage (pIII kapena pVIII). Kuphatikizika kwa jini, Ndi kuwonetsedwa pamwamba pa bacteriophages. Pakatikati paukadaulo uwu ndikumanga laibulale yowonetsera phage, yomwe ili ndi mwayi kuposa malaibulale achilengedwe chifukwa imatha kukhala ndi zomangira zenizeni. Pambuyo pake, ma antibodies okhala ndi ma antigen apadera amawunikiridwa kudzera munjira yosankha zamoyo, ma antigen omwe amawatsata amakhazikika, ma phage osamangika amatsukidwa mobwerezabwereza, ndipo ma phage omangika amatsukidwa kuti alemeredwenso. Pambuyo pa maulendo atatu kapena kuposerapo kubwerezabwereza, kutsimikizika kwakukulu ndi ma antibodies ogwirizana kwambiri amasiyanitsidwa.

Chithunzi 3: Kumanga Library ya Antibody ndi Kuwunika

Ukadaulo wa recombinant DNA ungagwiritsidwe ntchito kupanga tizidutswa ta ma antibody. Ma antibodies a Fab amatha kupangidwa ndi hydrolyzed ndi chapamimba protease kupanga (Fab ') 2 zidutswa, zomwe zimagayidwa ndi papain kuti apange zidutswa za Fab. Chidutswa cha Fv chimakhala ndi VH ndi VL, zomwe zimakhala zosakhazikika bwino chifukwa chosowa ma disulfide bond. Choncho, VH ndi VL zimagwirizanitsidwa pamodzi kudzera mu peptide yaifupi ya 15-20 amino acid kuti ipange kachidutswa kakang'ono (scFv) antibody yokhala ndi molecular yolemera pafupifupi 25KDa.

Kufufuza kwa ma antibody ku Camelidae (Ngamila, LIama, ndi Alpaca) kwawonetsa kuti ma antibodies ali ndi maunyolo olemera okha komanso opanda unyolo wopepuka, motero amatchedwa ma antibodies olemera (hcAb). Malo osinthika a ma antibodies olemera amatchedwa single domain antibodies kapena nanobodies kapena VHH, yokhala ndi kukula kwa 12-15 kDa. Monga ma monomers, alibe zomangira za disulfide ndipo ndi okhazikika kwambiri, okhala ndi kuyanjana kwakukulu kwa ma antigen.

Chithunzi 5: Ma Antibody Olemera ndi VHH / Nanobody

Kulankhula kwa ma cell kumagwiritsa ntchito mawu akuti DNA yachilengedwe kapena yopangidwa kuti ikwaniritse kaphatikizidwe ka mapuloteni mu vitro, makamaka pogwiritsa ntchito E. coli expression system. Amapanga mapuloteni mwachangu ndikupewa kulemedwa kwa kagayidwe kachakudya ndi cytotoxic pama cell popanga mapuloteni ambiri ophatikizanso mu vivo. Ikhozanso kupanga mapuloteni ovuta kupanga, monga omwe amavuta kusintha pambuyo pomasulira kapena kupanga mapuloteni a membrane.