Antibodies, nanone bita immunoglobuline (Ig), ni glycoproteine ​​ihuza cyane na antigene.
 
Gutegura antibody bisanzwe bikorwa mugukingira inyamaswa no gukusanya antiserum.Kubwibyo, antiserum mubisanzwe irimo antibodies zirwanya izindi antigene zidafitanye isano nibindi bice bya poroteyine muri serumu.Muri rusange molekile ya antigen irimo epitopi nyinshi zitandukanye, antibodiyite zisanzwe nazo ni uruvange rwa antibodies zirwanya epitopi zitandukanye.Ndetse na antibodiyumu zisanzwe zirwanya epitope imwe ziracyagizwe na antibodiyite za heterogeneous zakozwe na clone zitandukanye B.Kubwibyo, antibodiyumu zisanzwe zitwa nanone antibodiyite ya polyclone, cyangwa antibodiyite ya polyclone mugihe gito.
 
Antibody ya Monoclonal (antibody ya monoclonal) ni antibody ihwanye cyane ikorwa na clone imwe ya selile B kandi ikayobora gusa epitope yihariye.Ubusanzwe itegurwa nubuhanga bwa Hybridoma-tekinoroji ya antibody ya Hybridoma ishingiye ku buhanga bwa fusion selile, ihuza selile B nubushobozi bwo gusohora antibodies zihariye na selile myeloma hamwe nubushobozi butagira akagero muri B-selile ya Hybridoma.Iyi selile ya hybridoma ifite ibiranga selile yababyeyi.Irashobora kwiyongera mu gihe kitazwi kandi idapfa muri vitro nka selile myeloma, kandi irashobora guhuza no gusohora antibodi zihariye nka lymphocytes splenic.Binyuze muri cloni, umurongo wa monoclonal ukomoka kuri selile imwe ya hybridoma, ni ukuvuga umurongo wa Hybridoma, urashobora kuboneka.Antibodiyite ikora ni antibodiyite nyinshi zirwanya antigenic determinant, ni ukuvuga antibodiyite za monoclonal.
 
Antibodies zibaho nka monomers imwe cyangwa nyinshi Y (ni ukuvuga antibodiyite ya monoclonal cyangwa antibodiyite ya polyclone).Buri monomer Y igizwe na 4 polypeptide iminyururu, harimo iminyururu ibiri iremereye hamwe n'iminyururu ibiri imwe.Urunigi rwumucyo nu munyururu uremereye ukurikije uburemere bwa molekile.Hejuru yimiterere Y-imiterere ni akarere gahinduka, aribwo antigen ihuza urubuga.(Amagambo yavuye muri Detai Bio-Monoclonal Antibody Concepts)
 
Imiterere ya Antibody
Urunigi rukomeye
Hariho ubwoko butanu bwinyamabere Ig iminyururu iremereye, yitiriwe inyuguti zikigereki α, δ, ε, γ, na μ.Antibodies zihuye zitwa IgA, IgD, IgE, IgG, na IgM.Iminyururu iremereye itandukanye mubunini no mubigize.α na γ zirimo aside amine zigera kuri 450, mugihe μ na ε zirimo aside amine hafi 550.
Buri munyururu uremereye ufite uturere tubiri: akarere gahoraho nakarere gahinduka.Antibodies zose zubwoko bumwe zifite akarere gahoraho, ariko hariho itandukaniro hagati ya antibodies zubwoko butandukanye.Uturere duhoraho twumunyururu uremereye γ, α, na δ bigizwe na domaine eshatu Ig murwego rumwe, hamwe nakarere ka hinge kugirango byongere ubworoherane;uturere duhoraho twumunyururu uremereye μ na ε bigizwe na 4 Ig domaine.Agace gahindagurika k'urunigi ruremereye rwa antibody ikorwa na selile B zitandukanye ziratandukanye, ariko akarere gahindagurika ka antibody ikorwa na selile B imwe cyangwa clone selile nimwe, kandi akarere gahinduka kuri buri munyururu uremereye ni acide amine 110 z'uburebure., Kandi ukore Igikoresho kimwe.
 
Urunigi
Hariho ubwoko bubiri gusa bwiminyururu yoroheje yinyamabere: ubwoko bwa lambda nubwoko bwa kappa.Buri mucyo urumuri rufite domaine ebyiri zahujwe: akarere gahoraho nakarere gahinduka.Uburebure bwurumuri rwumucyo ni 211 ~ 217 acide amino.Iminyururu ibiri yoroheje iri muri buri antibody ihora ari imwe.Ku nyamaswa z’inyamabere, urunigi rw'urumuri muri buri antibody rufite ubwoko bumwe gusa: kappa cyangwa lambda.Mu binyabuzima bimwe na bimwe byo hasi, nk'amafi ya karitsiye (amafi ya karitsiye) n'amafi yo mu magufa, ubundi bwoko bw'iminyururu yoroheje nk'ubwoko bwa iota (iota).
 
Ibice bya Fab na Fc
Igice cya Fc gishobora guhuzwa neza na enzymes cyangwa amarangi ya fluorescent kugirango yandike antibodies.Nigice aho antibody izunguruka ku isahani mugihe cya ELISA, kandi ni nigice aho antibody ya kabiri imenyekana kandi igahuzwa na immunoprecipitation, immunoblotting na immunohistochemie.Antibodies zirashobora gushiramo hydrolyz mubice bibiri F (ab) nigice kimwe cya Fc na enzymes za proteolyique nka papain, cyangwa zirashobora gucika mukarere ka hinge na pepsin hanyuma hydrolyz ikagabanywa mugice kimwe cya F (ab) nigice kimwe cya Fc.Ibice bya antibody ya IgG rimwe na rimwe ni ingirakamaro cyane.Bitewe no kubura igice cya Fc, igice cya F (ab) ntikizagwa hamwe na antigen, ntanubwo kizafatwa ningirabuzimafatizo z'umubiri mubushakashatsi bwa vivo.Kubera uduce duto twa molekuline no kubura imikorere ihuza (kubera kubura igice cya Fc), igice cya Fab gikunze gukoreshwa kuri radiolabeling mubushakashatsi bwimikorere, naho igice cya Fc gikoreshwa cyane nkibintu bibuza kwanduza amateka.
 
Uturere duhinduka kandi duhoraho
Agace gahinduka (akarere ka V) gaherereye kuri 1/5 cyangwa 1/4 (kirimo ibisigisigi bya aside amine bigera kuri 118) byumunyururu wa H hafi ya N-terminus na 1/2 (birimo ibisigisigi bya aside amine hafi 108-111) hafi ya N-terminus ya L.Buri karere ka V gafite impeta ya peptide ikozwe mumurongo wa disulfide, kandi buri mpeta ya peptide irimo ibisigisigi bya aside amine bigera kuri 67 kugeza 75.Ibigize hamwe na gahunda ya aside amine mukarere ka V bigena antigen ihuza umwihariko wa antibody.Bitewe nubwoko bugenda buhindagurika hamwe nurutonde rwa acide amine mukarere ka V, ubwoko bwinshi bwa antibodi hamwe na antigen zitandukanye zihuza zirashobora gushirwaho.Uturere V twumunyururu wa L na H bita VL na VH.Muri VL na VH, aside amine igizwe nurutonde rwuturere tumwe na tumwe dufite urwego rwo hejuru rwo gutandukana.Utu turere twitwa uturere twa hypervariable (HVR).Amino acide ya amino hamwe no gutondekanya ibice bitari HVR mukarere ka V birasa nkaho ari conservateur, aribyo bita akarere.Hariho uturere dutatu twa hypervariable muri VL, mubisanzwe biherereye mubisigisigi bya aside amine 24 kugeza 34 na 89 kugeza 97.HVR eshatu za VL na VH zitwa HVR1, HVR2 na HVR3.Ubushakashatsi nisesengura rya X-ray kristal diffaction byagaragaje ko akarere ka hypervariable koko ariho hantu antigen ya antibody ihuza, bityo ikitwa akarere kagena ubwuzuzanye (CDR).HVR1, HVR2 na HVR3 ya VL na VH birashobora kwitwa CDR1, CDR2 na CDR3.Mubisanzwe, CDR3 ifite urwego rwo hejuru rwa hypervariability.Agace ka hypervariable nacyo kibanza nyamukuru aho imitekerereze idasanzwe ya molekile ya Ig iba.Kenshi na kenshi, urunigi H rufite uruhare runini muguhuza antigen.
Agace gahoraho (C akarere)iherereye kuri 3/4 cyangwa 4/5 (hafi ya aside amine 119 kugeza kuri C terminal) yumurongo wa H hafi ya C na 1/2 (kirimo ibisigisigi bya aside amine bigera kuri 105) hafi ya C terminus ya L.Buri karere gakora k'urunigi H karimo ibisigazwa bya aside amine bigera ku 110, kandi birimo impeta ya peptide igizwe na 50-60 ya aside amine isigara ihujwe na disulfide.Aminide acide igizwe na gahunda yo muri kano karere birahoraho muburyo bumwe bwinyamanswa Ig isotype L hamwe nubwoko bumwe H.Kimwe, irashobora guhuza gusa na antigen ihuye, ariko imiterere yakarere kayo C ni imwe, ni ukuvuga ko ifite antigenicite imwe.Ifarashi irwanya abantu IgG ya kabiri ya antibody (cyangwa anti-antibody) irashobora guhuzwa hamwe na bibiri A guhuza antibodi (IgG) kurwanya exotoxine zitandukanye.Uru nirwo shingiro ryingenzi mugutegura antibodiyumu ya kabiri no gukoresha fluorescein, isotopes, enzymes nizindi antibodi zanditseho.
 
 
Ibicuruzwa bifitanye isano:
Akagari ka RT-qPCR kit