Serivisi yo Gukura Antibody

Yamazaki Inc.Irashobora gutanga antibody affinity yo gukura ukoresheje mutation na tekinoroji yo guhitamo. Mubisanzwe dukoresha scFv nkimiterere ya antibody mubikorwa bya serivise yo gukura kwa affinity hamwe na sisitemu yo kwerekana fagemide ya monovalent yakoreshejwe mu kugabanya ingaruka ziterwa no kwipimisha mugihe cyo gusuzuma antigen. Itsinda ryacu ryumwuga rirashobora gutanga serivisi zo gukura kuri antibodies imwe-imwe.

Yamazaki Inc.yishimiye gutanga serivisi zuzuye kugirango zihuze ibyifuzo bitandukanye byabakiriya bisi. Intego yacu nukwumva no guhuza ibyifuzo byabakiriya batandukanye no gufasha mubibazo byose biri imbere kandi bivuka mubikorwa byubushakashatsi.

Gukura kw'Ubusabane Niki?

Gukura kwa Affinity ni inzira antibodies zigenda zihindagurika kandi zigahitamo kugirango zongere umubano wa antigene yihariye. Ahanini biboneka muri germinal centre ya lymphhoide ya kabiri, selile B ikora hypermutation ya somatike no guhitamo bitewe na selile T na selile dendritic selile.
Mugihe cyo gukura kwa affinity, selile B ifite reseptor nkeya ya immunoglobuline yakirwa na antigen ihura kandi ikwirakwizwa no gutandukana. Binyuze muri somatike hypermutation affinity gukura, insimburangingo ya nucleotide idasanzwe iba mukarere kahinduye genes ya immunoglobuline, bigatuma habaho umusaruro wa antibody zitandukanye. Nyuma ya hypermutation ya somatike, guhitamo antigen ikorerwa kuri selile B hamwe na reseptor ya immunoglobuline. Utugingo ngengabuzima B dukora antibodies hamwe na antigenic yiyongera cyane cyane turagumya kandi twemerewe kwiyongera, mugihe selile B yagabanutse cyangwa idahindutse ihura na apoptose. Ubu buryo bwo gutondeka ihinduka no guhitamo biganisha ku gukora clone B selile, bikabyara antibodies zigenda ziyongera kuri antigen.

Serivisi yo Gukura Antibody

Ibyiza byingenzi byuburyo bushingiye kuri PCR ni uko ihinduka ryibanze ryibice byongerewe; igipimo cyamakosa kiroroshye kugenzura kandi uburyo bwihuse kandi bworoshye gushiraho kandi ntibukoresha imiti yangiza. Birazwi neza ko polymerase ya Taq ADN yigana ADN ubudahemuka buke, ahanini kubera ko nta gikorwa cyo gusuzuma 3 'kugeza 5'.Alpha Lifetech'santibody yubuhanga ikoresha uburyo bwa PCR bwibeshya kugirango ihindure cyane uturere twa CDR mugihe cyo kubaka amasomero. Kandi akenshi dukora ihinduka ryimiterere ihindagurika rwose mubice byose bya VH na VL, kugirango twongere ubwoko butandukanye bwibitabo byibitabo. Nyuma yo kubaka isomero rya antibody yahinduwe na PCR ikunze kwibeshya, gutoranya ibintu byinshi bihuza cyane (isano ya antibodies ya scFv irashobora kugera kuri 10 ^ 8 - 10 ^ 10 M) bikorwa no gutekesha igisubizo cyangwa kuri antigen itabujijwe hamwe no gukaraba byateganijwe neza kugirango hatorwe ibiciro.
  
Abahanga muriYamazaki Inc.koresha Escherichia coli mutator ihindura mutAD5TM nkimwe mubikorwa byinshi bya mutation kugirango utangire ihinduka ryimiterere, hanyuma uhindure isano no kwerekana ibice bya antibody recombinant. Imiterere yo gutoranya irashobora guhindurwa kubice bya antibody hamwe nu rwego rwo kongera umusaruro. Imiterere yo gukura muri selile ya Escherichia coli irashobora guhinduka kugirango hamenyekane ihinduka ryimiterere imwe kuri kilobase ya ADN, hafi ya kimwe na codon imwe kuri buri gice cya scFv. Nyuma yinzinguzingo nyinshi za mutation, kwerekana, no guhitamo, turashobora kunoza umubano uhoraho kuva 10 ^ 5 - 10 ^ 6 M kugeza 10 ^ 8 - 10 ^ 10 M.

Kunoza Ingamba zo Gukura

Imyanya imwe nimwe ya antibodies irashobora gutondekwa muburyo butandukanye bwasobanuwe (nka randomisation yuzuye hamwe na aside amine 20 yose cyangwa kubogama kubogamye hamwe na aside amine yatoranijwe ku ijanisha ryagenwe) kugirango bitezimbere. Dufite ingamba ebyiri zo kunoza antibody: icya mbere, ihinduka ryatangijwe kumyanya yabujijwe mukarere kagena ubwuzuzanye (CDRs) na site ya mutagenezi; icya kabiri, ihinduka ryinjizwa mu turere twose twa V-coding na mutagenez. Ukoresheje ubu buryo bubiri, urutonde rwa amino-acide yabitswe cyangwa akarere kose ka antibody izasimburwa nandi acide amine, ihujwe nubuhanga bwacu bwo gusikana ububiko bwibitabo bwa peptide, bishobora kongera cyane isano ya antibodi yinyungu kurwego rwo hejuru cyane.

Icyiciro Cyerekana-Antibody Afinity

Isomero rya scFv rya mutant rimaze kubakwa, ingamba ebyiri zo gusuzuma zirahari: biopanning hamwe na antigen ikomeye. Mubyambere, insimburangingo ya substrate ikoreshwa mugutandukanya antibody yo hejuru cyane, kubera ko ihinduka rito ryaba ryarogejwe, hasigara uduce twinshi twa fage. Ingamba ya kabiri ikoresha antigen yanditseho igisubizo, guhitamo gushingiye kuburinganire buhoraho (Kd), no guhitamo gushingiye kubihuza, ubu buryo bwo guhitamo butandukanya antibody hamwe na Kd nziza.

Niba ufite ikibazo, nyamuneka twandikire igihe icyo aricyo cyose.