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Muaj qhov sib txawv loj ntawm monoclonal thiab polyclonal antibodies. Cov kab mob monoclonal yog cov khaub ncaws ntau tshuaj tiv thaiv uas tau tsim los ntawm ib leeg B cell clone thiab lub hom phiaj tsuas yog qhov tshwj xeeb epitope. Hybridoma antibody thev naus laus zis yog tsim los ntawm kev siv thev naus laus zis ntawm tes, uas suav nrog kev nkag siab zoo B cells thiab myeloma cells nrog lub peev xwm ua kom tsis txawj tuag mus rau hauv B cell hybrids qog.
Ib leeg hybridoma ntawm tes nrog cov yam ntxwv no tau siv los cog cov pej xeem ntawm tes los npaj cov tshuaj tiv thaiv tshwj xeeb tiv thaiv epitope, uas yog, a monoclonal tshuaj tiv thaiv.
Kev tsim kho ntawm cov kab mob monoclonal tau hla plaub theem, uas yog:
Murine monoclonal tshuaj tiv thaiv, chimeric antibody, humanized monoclonal antibody thiab tag nrho tib neeg cov tshuaj tiv thaiv kab mob monoclonal.
Siab tib neeg cov kab mob monoclonal: Cheeb tsam sib txawv thiab thaj tsam tas li ntawm cov tshuaj tiv thaiv kab mob yog tib neeg keeb kwm, tshem tawm cov tshuaj tiv thaiv kab mob thiab cov tshuaj lom.
Cov thev naus laus zis cuam tshuam rau kev npaj ua tiav tib neeg cov tshuaj tiv thaiv feem ntau suav nrog:
Tib neeg siv tshuab thev naus laus zis, EBV hloov pauv B lymphocyte thev naus laus zis, phage zaub thev naus laus zis, transgenic nas tshuaj tiv thaiv kab mob npaj thev naus laus zis thiab tib leeg B cell tshuaj tiv thaiv kab mob.
Tib neeg thiab tag nrho humanized tshuaj tiv thaiv tshuaj npaj los ntawm tib neeg thiab muaj cov tshuaj tiv thaiv tib neeg muaj peev xwm kov yeej ntau yam tsis txaus ntawm cov tshuaj tiv thaiv tsiaj thiab cov tshuaj tiv thaiv chimeric vim tias lawv muaj kev sib raug zoo, tshwj xeeb, thiab muaj tshuaj lom tsawg thiab muaj kev phiv. Nws tau dhau los ua qhov tsis sib xws hauv kev txhim kho tshuaj kho tshuaj tiv thaiv kab mob.
Cov qhov dej zoo uas tau sim rau tshuaj tiv thaiv kab mob tuaj yeem tsim thiab cloned kom tau txais cov tshuaj tiv thaiv kab mob monoclonal zais los ntawm cov xeeb ntxwv ntawm ib lub cell. Feem ntau hais lus, ua ntej lub sijhawm cloning, zoo dua. Vim tias nyob rau lub sijhawm no, ntau yam hybridoma cells loj hlob zoo nyob rau tib lub sijhawm, sib tw rau kev noj zaub mov thiab chaw, thiab cov cell tsim cov tshuaj tiv thaiv tshwj xeeb tuaj yeem ua rau poob qis thiab tshem tawm.
Txawm li cas los xij, lub sijhawm cloning yuav tsum tsis txhob ntxov dhau, txwv tsis pub cov yam ntxwv ntawm tes tsis ruaj khov, thiab tus lej ntawm cov cell me me thiab poob yooj yim. Lub cloned positive hybridoma cells, tom qab lub sijhawm kev coj noj coj ua, tseem yuav plam lub peev xwm los tsim cov tshuaj tiv thaiv vim yog kev hloov pauv ntawm tes lossis poob ntawm cov chromosomes tshwj xeeb, yog li lawv yuav tsum tau cloned ntau thiab ntau dua. Tus naj npawb ntawm cloning yog txiav txim siab los ntawm lub peev xwm tso pa tawm thiab tiv thaiv kab mob ntawm antigens. Feem ntau hais lus, tus naj npawb ntawm clones ntawm lub zog tiv thaiv kab mob antigen tuaj yeem tsawg dua, tab sis tsawg kawg 3 txog 5 clones tuaj yeem ua rau nws ruaj khov. Muaj ntau txoj hauv kev ntawm cloning, suav nrog txwv txoj kev dilution, micromanipulation txoj kev, cov txheej txheem mos agar phaj thiab txoj kev ua kom sib cais ua kom pom kev zoo.
Txij li thaum lub advent ntawm cov kab mob monoclonal, lawv tau siv rau hauv ntau thaj chaw ntawm cov tshuaj vim lawv cov yam ntxwv tshwj xeeb.
Antigens feem ntau yog tsim los ntawm ntau qhov kev txiav txim siab antigenic. Ib hom kev txiav txim siab antigenic txhawb lub cev, thiab tshuaj tiv thaiv tsim los ntawm B lymphocyte tau txais cov tshuaj tiv thaiv kab mob hu ua Monoclonal Antibody. Lub cev tau txhawb nqa los ntawm ntau qhov kev txiav txim siab antigenic, thiab ntau yam tshuaj tiv thaiv kab mob monoclonal tau tsim raws li. Cov tshuaj tiv thaiv kab mob monoclonal no tau sib xyaw ua ke los tsim polyclonal tshuaj tiv thaiv.
Cov qauv txheej txheem ntawm cov tshuaj antigen uas tuaj yeem ua rau lub cev tsim cov tshuaj tiv thaiv hu ua epitope. Muaj peev xwm muaj ntau qhov sib txawv antigenic txiav txim siab ntawm antigen, yog li lub cev tsim ntau yam tshuaj tiv thaiv sib txawv, thiab cov tshuaj tiv thaiv zaum kawg yog cov ntshav ntshav. Cov pej xeem ntawm cov ntshav ntshav uas tsuas yog ua rau ntawm ib qho tshuaj tiv thaiv kab mob antigenic yog clone. Cov tshuaj tiv thaiv tshwj xeeb uas tsim los ntawm clone hu ua monoclonal antibody. Cov kab mob monoclonal tuaj yeem tshwj xeeb khi rau ib qho tshwj xeeb antigenic tus txiav txim siab, ib yam li lub foob pob hluav taws raug tsoo lub hom phiaj. Ntawm qhov tod tes, txawm hais tias nws yog tib qho kev txiav txim siab antigenic, ntau qhov clones tuaj yeem tsim cov tshuaj tiv thaiv kab mob hauv lub cev, tsim kev sib xyaw ntawm ntau cov tshuaj tiv thaiv monoclonal, hu ua polyclonal tshuaj tiv thaiv.
Antigens feem ntau yog tsim los ntawm ntau yam epitopes. Ib hom epitope txhawb lub cev. Cov tshuaj tiv thaiv kab mob tsim los ntawm B lymphocyte tau txais lub antigen hu ua monoclonal antibody. Lub cev tau txhawb los ntawm ntau yam kev txiav txim siab antigenic, thiab ntau yam tshuaj tiv thaiv kab mob monoclonal tau tsim raws li. Cov tshuaj tiv thaiv kab mob monoclonal no tau sib xyaw ua ke los tsim cov tshuaj tiv thaiv kab mob polyclonal. Tsuas yog ntau qhov kev txiav txim siab antigenic, tib hom epitope tseem tuaj yeem txhawb lub cev los tsim tsib hom tshuaj tiv thaiv kab mob: IgG, IgM, IgA, IgE thiab IgD.
Polyclonal antibodies yog ib pawg immunoglobulins zais los ntawm lub cev cov ntshav ntshav uas txhawb los ntawm heterologous antigens (macromolecular antigens, hapten conjugates). Cov tshuaj tiv thaiv kab mob Polyclonal tau siv dav hauv kev tshawb fawb thiab kuaj mob vim tias lawv muaj peev xwm paub ntau yam epitopes thiab ua rau muaj dej nag tshwm sim nrog lub sijhawm npaj luv thiab tus nqi qis.
Ib lub polyclonal antiserum zoo muaj ntau yam tshuaj tiv thaiv tiv thaiv epitopes sib txawv ntawm qee yam antigen. Txij li polyclonal antiserum feem ntau muaj cov tshuaj tiv thaiv tiv thaiv epitopes sib txawv ntawm qee yam antigen, suav nrog cov tsis muaj peev xwm tiv taus epitopes, nws kuj tseem yuav muaj qhov cuam tshuam hauv qhov piv txwv tob. Hauv kev ua kom tawv nqaij ntawm cov ntaub so ntswg ua ntu ntu, Polyclonal cov tshuaj tiv thaiv kab mob feem ntau siv. Raws li qhov xav tau sib txawv ntawm qhov kev sim, polyclonal antibodies tau siv los sau cov kab mob sib xws.
Ib qho ntxiv, hauv kev ua liaj ua teb, polyclonal tshuaj tiv thaiv yog siv rau kev saib xyuas ntawm qhov chaw ntawm cov tshuaj tua kab; hauv kev siv tshuaj kho mob, tshuaj tiv thaiv kab mob polyclonal feem ntau yog siv rau kev kuaj kab mob, kuaj mob thiab kho, xws li cov tshuaj tiv thaiv kab mob tiv thaiv kab mob rau kev hloov pauv thiab kho cov kab mob autoimmune.
Cloning: hais txog asexually reproduced cell line, uas yog pawg ntawm cov kab xov tooj ntawm tes tsim los ntawm kev faib thiab rov ua dua tshiab ntawm ib tus zuj zus ntawm tes. Hauv txhua tus tswv cuab ntawm tsev neeg no, yog tias tsis muaj kev hloov pauv, cov noob yog tib yam nkaus.
Polyclonal tshuaj tiv thaiv (pAb): Txhaj tshuaj tiv thaiv tsiaj nrog antigen uas muaj ntau yam epitopes, uas tuaj yeem txhawb ntau yam B cell clones los tsim cov tshuaj tiv thaiv sib txawv rau ntau yam epitopes. Cov tshuaj tiv thaiv kab mob uas tau txais yog qhov sib xyaw ntawm ntau yam tshuaj tiv thaiv.
Cov kab mob monoclonal (mAb): Cov tshuaj tiv thaiv kab mob homologous tsim los ntawm clone ntawm B cells uas paub txog epitope. Siab zoo ib yam, muaj zog tshwj xeeb, muaj lub npe siab, me me lossis tsis muaj kev cuam tshuam hla.
Qhov sib txawv ntawm kev npaj tshuaj tiv thaiv kab mob monoclonal:
B lymphocytes kho nrog cov antigens tshwj xeeb thiab myeloma cells tau fused kom tau txais cov cell hybridoma. Tom qab tshuaj xyuas los ntawm HAT nruab nrab thiab kuaj pom lub ntsiab lus los ntawm ELISA, tau txais cov clone zoo, thiab tom qab ntawd ua cov kab lis kev cai ntawm tes lossis txhaj cov cell mus rau hauv lub plab kab noj hniav ntawm cov tsiaj (feem ntau yog kab laug sab/c nas) thiab coj lawv nrog ascites, sau cov supernatant/ascites thiab ntxuav lawv kom tau txais cov kab mob monoclonal. .
Kev npaj ntawm polyclonal tshuaj tiv thaiv tsis yog qhov nyuaj li cov tshuaj tiv thaiv monoclonal. Koj tsuas yog yuav tsum tau txhaj tshuaj antigen (siab dua qhov ua kom huv, zoo dua) ncaj qha rau hauv lub cev tsiaj txhawm rau txhaj tshuaj tiv thaiv kab mob. Tom qab 3 txog 4 qhov kev txhaj tshuaj tiv thaiv kab mob, tus titer tau txais kev tsim nyog los ntawm ELISA, thiab cov ntshav tau muab tso rau hauv ib nrab. Tus supernatant tuaj yeem ua kom huv kom tau txais cov tshuaj tiv thaiv polyclonal. Yog li ntawd, kev npaj ua ntu zus ntawm cov tshuaj tiv thaiv kab mob polyclonal yog luv dua li cov tshuaj tiv thaiv kab mob monoclonal, thiab tus nqi pib npaj qis dua li ntawm cov tshuaj tiv thaiv monoclonal.
A. Monoclonal antibody
Zoo:
Zoo heev affinity thiab purification:
B. Polyclonal antibody
Zoo:
tsis zoo:
Cov yam ntxwv | Monoclonal Antibody | Polyclonal Antibody | |
---|---|---|---|
1 | Ntsig | muaj zog | Tsis muaj zog |
2 | stability | Tus neeg tsis zoo, rhiab rau lub cev thiab tshuaj lom neeg | Zoo |
3 | Yuav tsum tau txhaj tshuaj tiv thaiv kab mob | Cov tshuaj tiv thaiv tsis huv kuj tseem tuaj yeem tsim cov tshuaj tiv thaiv kab mob kom huv | Yuav tsum muaj cov tshuaj tiv thaiv kab mob siab zoo |
4 | standardization | Yooj yim rau tus qauv, qhov sib txawv me me ntawm pawg | Nyuaj, loj sib txawv nruab nrab ntawm pawg |
5 | Recognition | Tsuas yog ib qho epitope ntawm antigen raug kuaj pom | Muaj peev xwm paub ntau yam epitopes |
6 | Hla Kev Tawm Tsam | tsis yooj yim hla kev cuam tshuam nrog lwm cov protein | ib qho thiab nyuaj kom tsis txhob muaj kev cuam tshuam tshwj xeeb |
7 | Dej nag thiab agglutination | Feem ntau tsis | Common |
8 | Cov Nqi Ntau Lawm | High | Tsawg |
9 | Lub Sijhawm Npaj | ntev | luv luv |
10 | nqe | High | Tsawg |
Qhov sib txawv nruab nrab ntawm monoclonal thiab polyclonal antibodies yog qhov loj heev, thiab cov kev sib txawv no ua rau lawv muaj txiaj ntsig hauv cov haujlwm sib txawv. Cov kab mob monoclonal nyob hauv qhov chaw tseem ceeb hauv tshuaj, thaum polyclonal tshuaj tiv thaiv ci ntsa iab hauv thaj tsam ntawm kev ua liaj ua teb thiab nrhiav kab mob thiab kho mob.
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