Antibody Engineering yog dab tsi?

Nrog kev pab los ntawm antibody engineering, nws muaj peev xwm hloov kho qhov loj ntawm molecular, pharmacokinetics, immunogenicity, binding affinity, tshwj xeeb thiab effector muaj nuj nqi ntawm cov tshuaj tiv thaiv. Tom qab synthesizing cov tshuaj tiv thaiv, qhov tshwj xeeb khi ntawm cov tshuaj tiv thaiv ua rau lawv muaj txiaj ntsig zoo hauv kev kuaj mob thiab kev kho mob. Los ntawm kev siv tshuaj tiv thaiv kab mob, lawv tuaj yeem ua tau raws li cov kev xav tau ntawm cov tshuaj thiab kev kuaj mob thaum ntxov.

Lub hom phiaj ntawm kev siv tshuaj tiv thaiv kab mob yog tsim thiab tsim cov haujlwm tshwj xeeb, ruaj khov uas cov tshuaj tiv thaiv ntuj tsis tuaj yeem ua tiav, tso lub hauv paus rau kev tsim cov tshuaj tiv thaiv kab mob.

Alpha Lifetech, nrog rau nws qhov kev paub dav dav hauv kev tsim cov tshuaj tiv thaiv kab mob, tuaj yeem muab cov kev pabcuam monoclonal thiab polyclonal antibody rau ntau hom, nrog rau phage zaub antibody tsev qiv ntawv tsim kho thiab tshuaj xyuas cov kev pabcuam. Alpha Lifetech tuaj yeem muab cov neeg siv khoom zoo biosimilar antibodies thiab recombinant protein cov khoom, nrog rau cov kev pabcuam sib raug, tsim kom muaj txiaj ntsig zoo, tshwj xeeb, thiab ruaj khov tshuaj tiv thaiv. Los ntawm kev siv cov tshuaj tiv thaiv kab mob dav dav, cov txheej txheem protein thiab phage cov tshuab, peb muab cov kev pabcuam npog cov dej ntws thiab cov dej ntws tawm ntawm cov tshuaj tiv thaiv kab mob, suav nrog cov kev pabcuam xws li tshuaj tiv thaiv tib neeg, tshuaj tua kab mob purification, antibody sequencing, thiab antibody validation.

Lub pioneering theem ntawm antibody engineering muaj feem xyuam rau ob technologies:

--Recombinant DNA technology

--Hybridoma technology

Txoj kev loj hlob sai ntawm antibody engineering muaj feem xyuam nrog peb lub thev naus laus zis tseem ceeb:

--Gene cloning technology thiab polymerase saw cov tshuaj tiv thaiv

--Protein expression: Recombinant proteins yog tsim los ntawm cov txheej txheem kev qhia xws li cov poov xab, cov kab mob zoo li tus kab mob, thiab cov nroj tsuag

--Computer aided structural design

Thawj tiam ntawm cov tshuaj tiv thaiv yog humanized rau zus tau tej cov chimeric antibodies, qhov twg lub variable cheeb tsam ntawm nas monoclonal antibodies tau txuas mus rau thaj tsam tas li ntawm tib neeg IgG molecules. Lub antigen binding cheeb tsam (CDR) ntawm lub thib ob tiam nas monoclonal antibody tau hloov mus rau hauv tib neeg IgG. Tsuas yog rau cheeb tsam CDR, tag nrho lwm cov tshuaj tiv thaiv yog yuav luag tib neeg cov tshuaj tiv thaiv kab mob, thiab kev siv zog ua kom tsis txhob ua rau tib neeg cov tshuaj tiv thaiv nas antibody (HAMA) cov lus teb thaum siv nas clone antibodies rau tib neeg kev kho mob.

 

Txhawm rau tsim lub tsev qiv ntawv phage, thawj kauj ruam yog kom tau cov noob encoding cov tshuaj tiv thaiv kab mob, uas tuaj yeem raug cais tawm ntawm B hlwb ntawm cov tsiaj tiv thaiv kab mob (immune tsev qiv ntawv), muab rho tawm ncaj qha los ntawm cov tsiaj tsis muaj tshuaj tiv thaiv (natural library tsim), lossis txawm sib sau ua ke hauv vitro nrog cov tshuaj tiv thaiv kab mob (synthetic library). Tom qab ntawd, cov noob tau nthuav dav los ntawm PCR, muab tso rau hauv plasmids, thiab nthuav tawm hauv cov txheej txheem tsim nyog (poj niam qhia (feem ntau Pichia pastoris), prokaryotic qhia (feem ntau E. coli), mammalian cell qhia, cog cell qhia, thiab kab kab qhia kab mob kis kab mob rod-zoo li tus kab mob). Qhov ntau tshaj plaws yog E. coli qhia qhov system, uas sib koom ua ke ib qho tshwj xeeb encoding antibody ib ntu mus rau phage thiab encodes ib qho ntawm phage plhaub proteins (pIII lossis pVIII). Cov noob fusion ntawm, Thiab tso tawm rau ntawm qhov chaw ntawm bacteriophages. Lub hauv paus ntawm cov thev naus laus zis no yog tsim lub tsev qiv ntawv phage zaub, uas muaj qhov zoo dua li cov tsev qiv ntawv ntuj nyob rau hauv uas nws tuaj yeem muaj kev khi tshwj xeeb. Tom qab ntawd, cov tshuaj tiv thaiv kab mob nrog cov tshuaj tiv thaiv tshwj xeeb raug tshuaj xyuas los ntawm cov txheej txheem kev xaiv tshuaj lom neeg, cov hom phiaj antigens raug kho, cov phages tsis raug muab rov qab ntxuav dua, thiab cov phages khi tau muab ntxuav kom huv ntxiv. Tom qab peb lossis ntau qhov kev rov ua dua, qhov tshwj xeeb siab thiab siab affinity antibodies raug cais tawm.

Fig 3: Antibody Library Kev tsim kho thiab tshuaj ntsuam

Recombinant DNA thev naus laus zis tuaj yeem siv los tsim cov tshuaj tiv thaiv kab mob. Fab cov tshuaj tiv thaiv kab mob tuaj yeem pib tsuas yog hydrolyzed los ntawm gastric protease los tsim (Fab ') 2 fragments, uas tom qab ntawd zom los ntawm papain los tsim ib tus neeg Fab tawg. Fv fragment muaj VH thiab VL, uas muaj kev ruaj ntseg tsis zoo vim tsis muaj disulfide bonds. Yog li ntawd, VH thiab VL tau txuas ua ke los ntawm peptide luv luv ntawm 15-20 amino acids los tsim ib qho kev sib txawv ntawm cov kab sib txawv (scFv) cov tshuaj tiv thaiv nrog ib qho hnyav molecular ntawm kwv yees li 25KDa.

Kev kawm txog cov qauv tshuaj tiv thaiv kab mob hauv Camelidae (Camel, LIama, thiab Alpaca) tau qhia meej tias cov tshuaj tiv thaiv tsuas muaj cov saw hnyav thiab tsis muaj lub teeb chains, yog li lawv hu ua heavy chain antibodies (hcAb). Qhov sib txawv ntawm cov saw hnyav cov tshuaj tiv thaiv kab mob hu ua ib leeg tshuaj tiv thaiv kab mob lossis nanobodies lossis VHH, nrog qhov loj ntawm 12-15 kDa. Raws li monomers, lawv tsis muaj disulfide daim ntawv cog lus thiab ruaj khov heev, nrog rau kev sib raug zoo heev rau antigens.

Fig 5: Heavy Chain Antibody thiab VHH/Nanobody

Cell dawb qhia siv cov lus qhia ntawm ntuj lossis hluavtaws DNA kom ua tiav hauv vitro protein synthesis, feem ntau yog siv E. coli qhia system. Nws tsim cov proteins sai sai thiab zam lub nra ntawm cov metabolism thiab cytotoxic ntawm cov hlwb thaum tsim cov protein ntau recombinant hauv vivo. Nws kuj tseem tuaj yeem tsim cov proteins uas nyuaj rau kev sib txuas, xws li cov uas nyuaj rau kev hloov kho tom qab kev txhais lus lossis synthesize membrane proteins.