Kwayoyin rigakafi, wanda kuma ake kira immunoglobulins (Ig), sune glycoproteins waɗanda ke ɗaure musamman ga antigens.
 
Ana samar da shirye-shiryen rigakafi na al'ada ta hanyar rigakafi da dabbobi da tattara maganin rigakafi.Don haka, maganin antiserum yakan ƙunshi ƙwayoyin rigakafi da sauran antigens marasa alaƙa da sauran abubuwan furotin a cikin jini.Gabaɗaya kwayoyin antigen galibi suna ɗauke da epitopes daban-daban, don haka ƙwayoyin rigakafi na al'ada suma cakuda ƙwayoyin rigakafi ne akan epitopes daban-daban.Hatta ƙwayoyin rigakafin ƙwayoyin cuta na al'ada waɗanda aka yi wa maganin epitope iri ɗaya har yanzu sun ƙunshi ƙwayoyin rigakafi iri-iri waɗanda ke haifar da clones na ƙwayoyin B daban-daban.Don haka, ƙwayoyin rigakafi na al'ada kuma ana kiran su polyclonal antibodies, ko polyclonal antibodies a takaice.
 
Monoclonal antibody (monoclonal antibody) wani rigakafi ne na musamman wanda aka samar da kwayar halitta ta B guda ɗaya kuma ana jagorantar shi kawai akan takamaiman epitope.Yawancin lokaci ana shirya shi ta hanyar fasahar hybridoma - fasahar antibody hybridoma ta dogara ne akan fasahar fusion cell, hada ƙwayoyin B tare da ikon ɓoye takamaiman ƙwayoyin cuta da ƙwayoyin myeloma tare da ƙarfin haɓaka mara iyaka a cikin hybridomas B-cell.Wannan tantanin halitta hybridoma yana da sifofin tantanin halitta na iyaye.Yana iya yaduwa har abada kuma mara mutuwa a cikin vitro kamar ƙwayoyin myeloma, kuma yana iya haɗawa da ɓoye takamaiman ƙwayoyin cuta kamar ƙwayoyin lymphocytes na splenic.Ta hanyar cloning, ana iya samun layin monoclonal wanda aka samo daga tantanin halitta na hybridoma guda ɗaya, wato, layin salula na hybridoma.Kwayoyin rigakafin da ta ke samarwa su ne ƙwayoyin rigakafi masu kama da juna a kan ƙayyadaddun antigenic iri ɗaya, wato, ƙwayoyin rigakafi na monoclonal.
 
Kwayoyin rigakafi sun wanzu azaman monomers ɗaya ko fiye da Y (watau, ƙwayoyin rigakafi na monoclonal ko polyclonal antibodies).Kowane monomer mai siffar Y yana kunshe da sarƙoƙi na polypeptide guda 4, gami da sarƙoƙi masu nauyi iri ɗaya da sarƙoƙi masu haske iri ɗaya.Sarkar haske da sarka mai nauyi ana sunaye bisa ga nauyin kwayoyin su.saman tsarin Y-dimbin yawa shine yanki mai canzawa, wanda shine wurin daurin antigen.(An karbo daga Detai Bio-Monoclonal Antibody Concept)
 
Tsarin rigakafi
Sarkar nauyi
Akwai nau'ikan sarƙoƙi masu nauyi na mammalian Ig guda biyar, masu suna tare da haruffan Helenanci α, δ, ε, γ, da μ.Kwayoyin rigakafi masu dacewa ana kiran su IgA, IgD, IgE, IgG, da IgM.Daban-daban sarƙoƙi masu nauyi sun bambanta da girman da abun da ke ciki.α da γ sun ƙunshi kusan amino acid 450, yayin da μ da ε sun ƙunshi kusan amino acid 550.
Kowace sarkar mai nauyi tana da yankuna biyu: yanki na dindindin da yanki mai canzawa.Dukkanin ƙwayoyin rigakafi na nau'in iri ɗaya suna da yanki iri ɗaya, amma akwai bambance-bambance tsakanin ƙwayoyin rigakafi na nau'ikan daban-daban.Yankuna akai-akai na sarƙoƙi masu nauyi γ, α, da δ sun ƙunshi yanki na Ig guda uku a cikin tandem, tare da yankin hinge don haɓaka sassaucin sa;yankuna akai-akai na sarƙoƙi masu nauyi μ da ε sun ƙunshi yanki na 4 Ig.Matsalolin da ke tattare da nau'in nau'in nau'in antibody da ke samar da kwayoyin B daban-daban ya bambanta, amma madaidaicin yanki na antibody da aka samar da tantanin halitta B ko tantanin halitta iri daya ne, kuma madaidaicin yanki na kowane sarkar mai nauyi ya kai kimanin amino acid 110 a tsayi., Kuma kafa yankin Ig guda ɗaya.
 
Sarkar haske
Akwai nau'ikan sarƙoƙin haske guda biyu kawai a cikin dabbobi masu shayarwa: nau'in lambda da nau'in kappa.Kowace sarkar haske tana da alaƙa guda biyu masu alaƙa: yanki na dindindin da yanki mai canzawa.Tsawon sarkar haske kusan 211 ~ 217 amino acid.Sarƙoƙin haske guda biyu da ke cikin kowane antibody koyaushe iri ɗaya ne.Ga dabbobi masu shayarwa, sarkar haske a cikin kowane maganin rigakafi yana da nau'i ɗaya kawai: kappa ko lambda.A wasu ƙananan kashin baya, irin su kifin cartilaginous (fishes cartilage) da kifin ƙashi, ana kuma samun wasu nau'ikan sarƙoƙi na haske kamar nau'in iota (iota).
 
Fab da Fc sassan
Za'a iya haɗa ɓangaren Fc kai tsaye tare da enzymes ko rini mai kyalli don lakabin ƙwayoyin cuta.Shi ne bangaren da antibody rivets a kan faranti a lokacin aiwatar da ELISA, kuma shi ne kuma bangaren da na biyu antibody aka gane da kuma daure a immunoprecipitation, immunoblotting da immunohistochemistry.Ana iya sanya ƙwayoyin rigakafi zuwa sassa biyu na F (ab) da kuma Fc ɗaya ta hanyar enzymes masu kariya irin su papain, ko kuma za a iya karya su daga yankin hinge ta pepsin kuma a sanya su cikin F(ab) 2 kashi da Fc daya.Gutsutturar rigakafin IgG wani lokaci suna da amfani sosai.Saboda rashin sashin Fc, sashin F (ab) ba zai yi hazo tare da antigen ba, kuma kwayoyin rigakafi ba za su kama shi ba a cikin nazarin vivo.Saboda ƙananan ɓangarorin ƙwayoyin cuta da rashin aikin haɗin kai (saboda rashin Fc sashi), ana amfani da sashin Fab don yin lakabi na rediyo a cikin karatun aiki, kuma ana amfani da sashin Fc azaman wakili mai toshewa a cikin tabo na histochemical.
 
Yankuna masu canzawa kuma akai-akai
Yankin m (yankin V) yana a 1/5 ko 1/4 (wanda ya ƙunshi kusan ragowar amino acid 118) na sarkar H kusa da N-terminus da 1/2 (mai ɗauke da ragowar amino acid 108-111) kusa da N-terminus na sarkar L.Kowane yanki na V yana da zoben peptide da aka kafa ta hanyar haɗin sarkar disulfide, kuma kowane zoben peptide ya ƙunshi kusan ragowar amino acid 67 zuwa 75.Ƙirƙiri da tsari na amino acid a cikin yankin V sun ƙayyade ƙayyadaddun dauri na antigen.Saboda sauye-sauyen nau'ikan amino acid da jerin amino acid a cikin yankin V, ana iya samar da nau'ikan rigakafi da yawa tare da takamaiman takamaiman antigen.Yankunan V na sarkar L da H ana kiran su VL da VH, bi da bi.A cikin VL da VH, abun da ke tattare da amino acid da jerin wasu yankuna na gida suna da matsayi mafi girma na bambancin.Ana kiran waɗannan yankuna hypervariable yankuna (HVR).Haɗin amino acid da tsari na sassan da ba HVR ba a cikin yankin V suna da ɗan ra'ayin mazan jiya, wanda ake kira yankin tsarin.Akwai yankuna uku masu saurin canzawa a cikin VL, yawanci suna samuwa a ragowar amino acid 24 zuwa 34 da 89 zuwa 97 bi da bi.HVR guda uku na VL da VH ana kiran su HVR1, HVR2 da HVR3, bi da bi.Bincike da nazarin diffraction na X-ray crystal sun tabbatar da cewa yankin hypervariable hakika shine wurin da antigen antigen ke ɗaure, don haka ana kiran shi yanki mai ƙayyadaddun bayanai (CDR).Ana iya kiran HVR1, HVR2 da HVR3 na VL da VH CDR1, CDR2 da CDR3 bi da bi.Gabaɗaya, CDR3 yana da matsayi mafi girma na haɓakawa.Yankin da ake iya canzawa kuma shine babban wurin da abubuwan da ke tantance kwayoyin Ig suka wanzu.A mafi yawan lokuta, sarkar H tana taka muhimmiyar rawa wajen ɗaure antigen.
Yankin dindindin (yankin C)yana a 3/4 ko 4/5 (kimanin daga amino acid 119 zuwa tashar C) na sarkar H kusa da tashar C da 1/2 (ya ƙunshi kusan ragowar amino acid 105) kusa da tashar C na sarkar L.Kowane yanki mai aiki na sarkar H yana ƙunshe da ragowar amino acid kusan 110, kuma ya ƙunshi zoben peptide wanda ya ƙunshi ragowar amino acid 50-60 waɗanda aka haɗa ta hanyar haɗin disulfide.Haɗin amino acid da tsarin wannan yanki sun dawwama a cikin dabba iri ɗaya Ig isotype L sarkar da nau'in H iri ɗaya.Hakanan, yana iya ɗaure musamman ga antigen ɗin da ya dace, amma tsarin yankin C iri ɗaya ne, wato yana da antigenicity iri ɗaya.Doki anti-human IgG secondary antibody (ko anti-antibody) za a iya hade tare da biyu A hade antibodies (IgG) da daban-daban exotoxins faruwa.Wannan muhimmin tushe ne don shirya ƙwayoyin rigakafi na biyu da yin amfani da fluorescein, isotopes, enzymes da sauran alamun rigakafin.
 
 
Samfura masu dangantaka:
Cell Direct RT-qPCR kit