Antibodies, tseem hu ua immunoglobulins (Ig), yog glycoproteins uas tshwj xeeb khi rau antigens.
 
Kev npaj tshuaj tiv thaiv kab mob yog tsim los ntawm kev txhaj tshuaj tiv thaiv tsiaj thiab sau cov tshuaj antiserum.Yog li ntawd, antiserum feem ntau muaj cov tshuaj tiv thaiv kab mob tiv thaiv lwm cov antigens tsis sib xws thiab lwm cov protein ntau hauv cov ntshav.General antigen molecules feem ntau muaj ntau qhov sib txawv epitopes, yog li cov tshuaj tiv thaiv kab mob kuj yog ib qho sib xyaw ntawm cov tshuaj tiv thaiv kab mob tiv thaiv ntau yam epitopes.Txawm tias cov tshuaj tiv thaiv kab mob sib xyaw ua ke tawm tsam tib lub epitope tseem muaj cov tshuaj tiv thaiv heterogeneous uas tsim los ntawm ntau hom B cell clones.Yog li ntawd, cov tshuaj tiv thaiv kab mob hauv cov ntshav tseem hu ua polyclonal antibodies, lossis polyclonal antibodies luv luv.
 
Monoclonal antibody (monoclonal antibody) yog ib qho tshuaj tiv thaiv zoo heev uas tsim los ntawm ib leeg B cell clone thiab tsuas yog qhia tawm tsam ib qho epitope.Nws feem ntau yog npaj los ntawm hybridoma technology-hybridoma antibody technology yog raws li cell fusion technology, combining B hlwb nrog lub peev xwm los secrete tej antibodies thiab myeloma hlwb nrog infinite loj hlob muaj peev xwm mus rau hauv B-cell hybridomas.Lub hybridoma cell no muaj cov yam ntxwv ntawm niam txiv cell.Nws muaj peev xwm proliferate indefinitely thiab immortally nyob rau hauv vitro zoo li myeloma hlwb, thiab nws muaj peev xwm synthesize thiab secrete tej antibodies zoo li splenic lymphocytes.Los ntawm cloning, ib kab monoclonal muab los ntawm ib leeg hybridoma cell, uas yog, hybridoma cell kab, tuaj yeem tau.Cov tshuaj tiv thaiv kab mob uas nws tsim muaj ntau hom tshuaj tiv thaiv kab mob tiv thaiv tib yam antigenic determinant, uas yog, monoclonal antibodies.
 
Cov tshuaj tiv thaiv muaj nyob ua ib lossis ntau dua Y-zoo li monomers (piv txwv li, monoclonal antibodies lossis polyclonal antibodies).Txhua Y-shaped monomer yog tsim los ntawm 4 polypeptide chains, suav nrog ob txoj hlua hnyav zoo ib yam thiab ob lub teeb sib xws.Lub teeb saw thiab cov saw hnyav muaj npe raws li lawv qhov hnyav molecular.Sab saum toj ntawm Y-zoo li tus qauv yog thaj tsam sib txawv, uas yog qhov chaw sib txuas antigen.(Excerpt los ntawm Detai Bio-Monoclonal Antibody Concept)
 
Antibody qauv
Hnyav saw
Muaj tsib hom tsiaj nyeg Ig hnyav chains, muaj npe nrog cov ntawv Greek α, δ, ε, γ, thiab μ.Cov tshuaj tiv thaiv uas raug hu ua IgA, IgD, IgE, IgG, thiab IgM.Cov saw hnyav sib txawv txawv ntawm qhov loj thiab muaj pes tsawg leeg.α thiab γ muaj kwv yees li 450 amino acids, thaum μ thiab ε muaj kwv yees li 550 amino acids.
Txhua lub saw hnyav muaj ob thaj tsam: thaj chaw tas li thiab thaj chaw sib txawv.Tag nrho cov tshuaj tiv thaiv ntawm tib hom muaj tib thaj chaw tas li, tab sis muaj qhov sib txawv ntawm cov tshuaj tiv thaiv ntawm ntau hom.Cov cheeb tsam tas li ntawm cov chains hnyav γ, α, thiab δ yog tsim los ntawm peb qhov Ig domains hauv tandem, nrog rau thaj tsam pob khawm kom nws yooj yim;cov cheeb tsam tas li ntawm cov chains hnyav μ thiab ε yog tsim los ntawm 4 Ig domains.Cov cheeb tsam sib txawv ntawm cov saw hnyav ntawm cov tshuaj tiv thaiv uas tsim los ntawm cov B hlwb sib txawv, tab sis thaj tsam sib txawv ntawm cov tshuaj tiv thaiv uas tsim los ntawm tib B cell lossis cell clone yog tib yam, thiab thaj tsam sib txawv ntawm txhua qhov hnyav yog li 110 amino acids ntev., Thiab tsim ib lub npe Ig.
 
Lub teeb saw
Tsuas muaj ob hom teeb pom kev hauv cov tsiaj txhu: hom lambda thiab hom kappa.Txhua lub teeb saw muaj ob qhov txuas txuas: thaj chaw tas li thiab thaj chaw sib txawv.Qhov ntev ntawm lub teeb saw yog hais txog 211 ~ 217 amino acids.Ob lub teeb sawb uas muaj nyob rau hauv txhua qhov tshuaj tiv thaiv ib txwm yog tib yam.Rau cov tsiaj nyeg, lub teeb saw hauv txhua qhov tshuaj tiv thaiv tsuas muaj ib hom: kappa lossis lambda.Hauv qee cov vertebrates qis dua, xws li cov ntses cartilaginous (cov ntses pob txha mos) thiab cov ntses pob txha, lwm hom teeb sawb xws li hom iota (iota) kuj pom.
 
Fab thiab Fc ntu
Fc ntu tuaj yeem txuas ncaj qha nrog enzymes lossis fluorescent dyes los sau cov tshuaj tiv thaiv.Nws yog ib feem uas cov tshuaj tiv thaiv kab mob rivets ntawm lub phaj thaum lub sij hawm ELISA txheej txheem, thiab nws kuj yog ib feem ntawm qhov thib ob antibody raug lees paub thiab khi rau hauv immunoprecipitation, immunoblotting thiab immunohistochemistry.Cov tshuaj tiv thaiv tuaj yeem raug hydrolyzed rau hauv ob ntu F (ab) thiab ib ntu Fc los ntawm cov enzymes proteolytic xws li papain, lossis lawv tuaj yeem tawg los ntawm pob khawm los ntawm pepsin thiab hydrolyzed rau hauv ib ntu F (ab) 2 thiab ib ntu Fc.IgG cov tshuaj tiv thaiv kab mob qee zaum muaj txiaj ntsig zoo.Vim qhov tsis muaj Fc ntu, ntu F (ab) yuav tsis tuaj yeem nrog cov antigen, thiab nws yuav tsis raug ntes los ntawm lub cev tiv thaiv kab mob hauv kev tshawb fawb vivo.Vim tias qhov me me me me thiab tsis muaj kev sib txuas ntawm kev sib txuas (vim tsis muaj Fc ntu), Fab ntu feem ntau yog siv rau radiolabeling hauv cov kev tshawb fawb ua haujlwm, thiab Fc ntu feem ntau yog siv los ua tus neeg saib xyuas hauv histochemical staining.
 
Cov cheeb tsam sib txawv thiab tsis tu ncua
Cov cheeb tsam sib txawv (V cheeb tsam) nyob ntawm 1/5 lossis 1/4 (muaj txog 118 amino acid residues) ntawm H saw ze rau N-terminus thiab 1/2 (muaj txog 108-111 amino acid residues) ze ntawm N-terminus ntawm L saw.Txhua cheeb tsam V muaj lub nplhaib peptide tsim los ntawm cov saw hlau disulfide, thiab txhua lub nplhaib peptide muaj kwv yees li 67 txog 75 amino acid residues.Cov muaj pes tsawg leeg thiab kev npaj ntawm cov amino acids nyob rau hauv cheeb tsam V txiav txim lub antigen khi tshwj xeeb ntawm cov tshuaj tiv thaiv.Vim yog qhov hloov pauv txhua yam thiab cov kab ke ntawm cov amino acids hauv cheeb tsam V, ntau hom tshuaj tiv thaiv kab mob sib txawv tuaj yeem tsim tau.Cov cheeb tsam V ntawm L saw thiab H saw yog hu ua VL thiab VH, feem.Hauv VL thiab VH, cov amino acid muaj pes tsawg leeg thiab ib ntus ntawm qee thaj chaw hauv cheeb tsam muaj ntau qhov sib txawv.Cov cheeb tsam no hu ua hypervariable regions (HVR).Cov amino acid muaj pes tsawg leeg thiab kev npaj ntawm cov tsis-HVR qhov chaw nyob rau hauv V cheeb tsam yog conservative, uas yog hu ua lub moj khaum cheeb tsam.Muaj peb thaj chaw sib txawv hauv VL, feem ntau nyob ntawm cov amino acid residues 24 txog 34 thiab 89 txog 97 feem.Peb HVRs ntawm VL thiab VH yog hu ua HVR1, HVR2 thiab HVR3, ntsig txog.Kev tshawb fawb thiab kev tsom xam ntawm X-ray siv lead ua diffraction tau ua pov thawj tias thaj av sib txawv yog qhov tseeb qhov chaw uas cov tshuaj tiv thaiv kab mob antigen khi, yog li nws yog hu ua thaj tsam kev txiav txim siab ntxiv (CDR).Lub HVR1, HVR2 thiab HVR3 ntawm VL thiab VH tuaj yeem hu ua CDR1, CDR2 thiab CDR3 raws.Feem ntau, CDR3 muaj qib siab dua ntawm qhov sib txawv.Lub cheeb tsam hypervariable kuj yog qhov chaw tseem ceeb uas muaj cov idiotypic determinants ntawm Ig molecules nyob.Feem ntau, H chain plays lub luag haujlwm tseem ceeb hauv kev khi rau cov antigen.
Thaj tsam tas mus li (C cheeb tsam)yog nyob ntawm 3/4 lossis 4/5 (kwv yees los ntawm amino acid 119 mus rau C davhlau ya nyob twg) ntawm H saw ze rau C terminus thiab 1/2 (muaj txog 105 amino acid residues) ze rau C terminus ntawm L saw.Txhua thaj chaw ua haujlwm ntawm H saw muaj txog 110 amino acid residues, thiab muaj lub nplhaib peptide tsim los ntawm 50-60 amino acid residues txuas nrog disulfide bonds.Cov amino acid muaj pes tsawg leeg thiab kev npaj ntawm thaj av no yog qhov tsis tu ncua ntawm tib tsiaj Ig isotype L saw thiab tib hom H saw.Ib yam, nws tsuas tuaj yeem khi rau qhov sib thooj antigen, tab sis cov qauv ntawm nws cheeb tsam C yog tib yam, uas yog, nws muaj tib yam antigenicity.Tus nees tiv thaiv tib neeg IgG thib ob cov tshuaj tiv thaiv kab mob (lossis tshuaj tiv thaiv kab mob) tuaj yeem ua ke nrog ob qho kev sib xyaw ntawm cov tshuaj tiv thaiv kab mob (IgG) tawm tsam cov exotoxins sib txawv tshwm sim.Qhov no yog lub hauv paus tseem ceeb rau kev npaj cov tshuaj tiv thaiv kab mob thib ob thiab siv fluorescein, isotopes, enzymes thiab lwm yam tshuaj tiv thaiv kab mob.
 
 
Yam khoom:
Cell Direct RT-qPCR cov khoom siv