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Ama-antibodies, abizwa nangokuthi ama-immunoglobulins (Ig), angama-glycoprotein abopha ngokuqondile kuma-antigen.
 
Ukulungiswa okujwayelekile kwama-antibody kukhiqizwa ngokugoma izilwane nokuqoqa i-antiserum.Ngakho-ke, i-antiserum ngokuvamile iqukethe amasosha omzimba ngokumelene namanye ama-antigen angahlobene nezinye izingxenye zamaprotheni ku-serum.Amangqamuzana avamile e-antigen ngokuvamile aqukethe ama-epitopes amaningi ahlukene, ngakho amasosha omzimba avamile ayingxube yamasosha omzimba ngokumelene nama-epitopes amaningi ahlukene.Ngisho namasosha omzimba avamile e-serum aqondiswe ku-epitope efanayo asakhiwa amasosha omzimba ahlukahlukene akhiqizwa ama-clone B cell ahlukene.Ngakho-ke, amasosha omzimba avamile e-serum abizwa nangokuthi amasosha omzimba e-polyclonal, noma amasosha omzimba e-polyclonal ngamafuphi.
 
I-Monoclonal antibody (i-monoclonal antibody) iyi-antibody efanayo kakhulu ekhiqizwa i-clone eyodwa ye-B cell futhi iqondiswe kuphela ngokumelene ne-epitope ethile.Ivamise ukulungiselelwa ubuchwepheshe be-hybridoma—ubuchwepheshe be-hybridoma antibody busekelwe kubuchwepheshe bokuhlanganiswa kwamaseli, okuhlanganisa amaseli B anekhono lokukhiqiza amasosha omzimba athile namaseli e-myeloma anamandla okukhula angapheli abe ama-B-cell hybridomas.Leli seli le-hybridoma linezici zeseli lomzali.Ingakhula ngokungapheli futhi ingafi ku-vitro njengamaseli e-myeloma, futhi ingahlanganisa futhi ikhiphe amasosha omzimba athile njengama-splenic lymphocyte.Ngokusebenzisa i-cloning, umugqa we-monoclonal otholakala kuseli elilodwa le-hybridoma, okungukuthi, umugqa weseli we-hybridoma, ungatholakala.Amasosha omzimba ewakhiqizayo angamasosha omzimba ahlobene kakhulu ngokumelene ne-antigenic determinant, okungukuthi, amasosha omzimba e-monoclonal.
 
Ama-antibody akhona njenge-monomer eyodwa noma amaningi amise okuka-Y (okungukuthi, amasosha omzimba e-monoclonal noma amasosha omzimba e-polyclonal).I-monomer ngayinye emise okuka-Y yakhiwe ngamaketango angu-4 e-polypeptide, afaka amaketango amabili asindayo afanayo namaketango amabili okukhanya afanayo.Iketango elilula neketango elisindayo liqanjwe ngokwesisindo sazo samangqamuzana.Ingaphezulu lesakhiwo esimise okwe-Y yisifunda esiguquguqukayo, okuyisizinda sokubopha i-antigen.(Kucashunwe ku-Detai Bio-Monoclonal Antibody Concept)
 
Isakhiwo se-antibody
1Iketanga elisindayo
Kunezinhlobo ezinhlanu zamaketanga asindayo ezincelisayo ze-Ig, eziqanjwe ngezinhlamvu zesiGreki u-α, δ, ε, γ, kanye no-μ.Amasosha omzimba ahambisanayo abizwa nge-IgA, IgD, IgE, IgG, kanye ne-IgM.Amaketanga asindayo ahlukene ayahluka ngobukhulu kanye nokwakheka.U-α no-γ aqukethe cishe ama-amino acid angama-450, kuyilapho u-μ no-ε aqukethe cishe ama-amino acid angama-550.
Iketango ngalinye elisindayo linezifunda ezimbili: isifunda esingashintshi kanye nesifunda esiguquguqukayo.Wonke amasosha omzimba ohlobo olufanayo anendawo efanayo engaguquki, kodwa kunomehluko phakathi kwamasosha omzimba ezinhlobo ezahlukene.Izifunda ezingaguquki zamaketango asindayo u-γ, α, kanye no-δ zakhiwe izizinda ezintathu ze-Ig ngokuhambisana, nesifunda sehinge ukuze kwandiswe ukuguquguquka kwayo;izifunda ezingaguquki zamaketanga asindayo u-μ no-ε zakhiwe izizinda ze-4 Ig.Isifunda esiguquguqukayo sochungechunge olusindayo lwe-antibody ekhiqizwa amaseli e-B ahlukene sihlukile, kodwa indawo eguquguqukayo ye-antibody ekhiqizwa yi-B cell efanayo noma i-cell clone iyafana, futhi indawo eguquguqukayo yeketango elisindayo ngalinye lilinganiselwa ku-110 amino acid ubude., Futhi yakha isizinda esisodwa se-Ig.
 
Iketango elikhanyayo
Kunezinhlobo ezimbili kuphela zamaketanga okukhanya ezilwaneni ezincelisayo: uhlobo lwe-lambda nohlobo lwe-kappa.Iketango ngalinye lokukhanya linezizinda ezimbili ezixhumene: isifunda esingashintshi kanye nesifunda esiguquguqukayo.Ubude beketango lokukhanya cishe bungama-211 ~ 217 amino acid.Amaketanga okukhanya amabili aqukethwe ku-antibody ngayinye ahlala efana.Ezilwaneni ezincelisayo, iketango lokukhanya ku-antibody ngayinye linohlobo olulodwa kuphela: i-kappa noma i-lambda.Kwezinye izilwane ezinomgogodla eziphansi, njengezinhlanzi ze-cartilaginous (izinhlanzi ze-cartilage) nezinhlanzi ezinamathambo, ezinye izinhlobo zamaketanga alula njengohlobo lwe-iota (iota) nazo ziyatholakala.
 
Izingxenye ze-Fab ne-Fc
Ingxenye ye-Fc ingahlanganiswa ngokuqondile nama-enzyme noma odayi be-fluorescent ukuze ilebula amasosha omzimba.Kuyingxenye lapho i-antibody igoba khona epuletini ngesikhathi senqubo ye-ELISA, futhi futhi iyingxenye lapho i-antibody yesibili ibonwa futhi iboshelwe ekuvimbeleni ukuzivikela komzimba, i-immunoblotting kanye ne-immunohistochemistry.Ama-antibody angenziwa i-hydrolyzed abe amasegimenti amabili ka-F(ab) kanye nesegmenti eyodwa ye-Fc ngama-enzyme e-proteolytic afana ne-papain, noma angephulwa endaweni eyihinge nge-pepsin futhi afakwe nge-hydrolyzed abe yisegimenti engu-F(ab)2 kanye nengxenye eyodwa ye-Fc.Izingcezu ze-IgG antibody kwesinye isikhathi ziwusizo kakhulu.Ngenxa yokushoda kwengxenye ye-Fc, ingxenye ye-F(ab) ngeke ibe ne-antigen, futhi ngeke ithathwe amaseli omzimba ezifundweni ze-vivo.Ngenxa yezingcezu ezincane zamangqamuzana kanye nokuntuleka komsebenzi wokuxhumanisa (ngenxa yokuntuleka kwengxenye ye-Fc), ingxenye ye-Fab ivame ukusetshenziselwa i-radiolabeling ezifundweni zokusebenza, futhi ingxenye ye-Fc isetshenziswa kakhulu njenge-ejenti evimbelayo ekungcoleni kwe-histochemical.
 
Izifunda eziguquguqukayo neziqhubekayo
Isifunda esiguquguqukayo (isifunda esingu-V) sitholakala ku-1/5 noma 1/4 (equkethe cishe izinsalela ze-amino acid eziyi-118) yeketango H eduze ne-N-terminus kanye no-1/2 (equkethe cishe izinsalela ze-amino acid ezingaba ngu-108-111) eduze kwe-N-terminus yeketango L.Isifunda ngasinye esingu-V sinendandatho ye-peptide eyakhiwe ngamabhondi e-intra-chain disulfide, futhi iringi ngayinye ye-peptide iqukethe cishe izinsalela ze-amino acid ezingama-67 kuya kwezingama-75.Ukwakheka nokuhlelwa kwama-amino acid endaweni ye-V kunquma ukucaciswa kokubopha kwe-antigen kwe-antibody.Ngenxa yezinhlobo ezihlala zishintsha kanye nokulandelana kwama-amino acid endaweni ye-V, izinhlobo eziningi zamasosha omzimba anemininingwane ehlukene ye-antigen ebophayo zingakhiwa.Izifunda ezingu-V zochungechunge lwe-L kanye ne-H zibizwa ngokuthi i-VL ne-VH, ngokulandelanayo.Ku-VL ne-VH, ukwakheka kwe-amino acid nokulandelana kwezinye izifunda zendawo kunezinga eliphakeme lokuhlukahluka.Lezi zifunda zibizwa nge-hypervariable regions (HVR).Ukwakheka kwe-amino acid nokuhlelwa kwezingxenye ezingezona eze-HVR esifundeni se-V zinomsoco uma kuqhathaniswa, okubizwa ngokuthi isifunda sohlaka.Kunezifunda ezintathu ze-hypervariable ku-VL, ngokuvamile ezitholakala ezinsalela ze-amino acid 24 kuya ku-34 kanye no-89 kuya ku-97 ngokulandelanayo.Ama-HVR amathathu e-VL ne-VH abizwa nge-HVR1, i-HVR2 ne-HVR3, ngokulandelana.Ucwaningo nokuhlaziywa kwe-X-ray crystal diffraction kufakazele ukuthi isifunda esingaguquguquki ngempela yindawo lapho i-antigen ibopha khona, ngakho ibizwa nge-complementarity-determining region (CDR).I-HVR1, i-HVR2 ne-HVR3 ye-VL ne-VH ingabizwa nge-CDR1, i-CDR2 ne-CDR3 ngokulandelanayo.Ngokuvamile, i-CDR3 inezinga eliphezulu lokungaguquguquki.Isifunda esingaguquguquki futhi siyindawo eyinhloko lapho izinqumo ze-idiotypic zama-molecule e-Ig zikhona.Ezimweni eziningi, uchungechunge lwe-H ludlala indima ebaluleke kakhulu ekubopheni i-antigen.
2Isifunda esingaguquki (isifunda C)itholakala ku-3/4 noma ku-4/5 (cishe isuka ku-amino acid 119 kuya ku-C terminal) yeketango elingu-H eduze kwetheminasi engu-C kanye no-1/2 (iqukethe cishe izinsalela ze-amino acid eziyi-105) eduze kwetheminasi engu-C yochungechunge lwe-L.Isifunda ngasinye esisebenzayo sochungechunge lwe-H siqukethe cishe izinsalela ze-amino acid eziyi-110, futhi iqukethe iringi ye-peptide eyakhiwe izinsalela ze-amino acid ezingama-50-60 ezixhunywe ngamabhondi e-disulfide.Ukwakheka nokuhlelwa kwe-amino acid yalesi sifunda akushintshile kuchungechunge lwesilwane esisodwa se-Ig isotype L kanye nohlobo olufanayo lweketango H.Ngokufanayo, ingakwazi ukubopha ngokuqondile i-antigen ehambisanayo, kodwa isakhiwo sesifunda sayo se-C siyafana, okungukuthi, sine-antigenicity efanayo.I-anti-human IgG antibody yesibili (noma i-anti-antibody) ingahlanganiswa nenhlanganisela emibili ye-A yamasosha omzimba (IgG) ngokumelene nama-exotoxin ahlukene eyenzekayo.Lesi yisisekelo esibalulekile sokulungiselela amasosha omzimba esibili nokusebenzisa i-fluorescein, isotopes, ama-enzyme namanye amasosha omzimba abhalwe phansi.
 
 
Imikhiqizo Ehlobene:
Ikhithi ye-Cell Direct RT-qPCR

 

 


Isikhathi sokuthumela: Sep-30-2021