Icyiciro cyo kwerekana Isomero ryubaka Serivisi

Alpha Lifetech amaze imyaka myinshi akora cyane mubuhanga bwo kwerekana Phage. Yubatse urubuga rwiza rwa tekinoroji ya Phage yerekana ikora neza kubushakashatsi bwubumenyi cyangwa ubushakashatsi bwumushinga kandi byoroshya umusaruro. Alpha Lifetech irashobora kandi guha abakiriya serivisi nka vhh antibody, umusaruro wa scFv antibody, umusaruro wa Fab Antibody.

Alpha Lifetech ifite urubuga rwuzuye rwa M13 / T7 rwo kwerekana ibyubaka no gutanga amasomero ya antibody. Turashobora kandi gutanga serivise yihariye, nkumusemburo wa antibody scFv hamwe no kugenzura antibody nyinshi, kugirango uhuze ibyo ukeneye.

Intangiriro kuri Phage yerekana

Ikoreshwa rya tekinoroji yerekana ni tekinike ikoresha ibice (virusi yanduza bagiteri) gushakisha molekile ikora ya poroteyine cyangwa peptide yihariye. Ubuhanga bwo kubaka isomero rya antibody harimo kubaka ububiko bwibitabo bwa Fab antibody, kubaka isomero rya antibody scFv, kubaka isomero rya nanobody, nibindi. Ukurikije ubwoko bwa bacteriofage, birashobora kugabanywa M13, T7, T4, λ, nubundi buryo. Ukurikije ubwoko bwibitabo, burashobora kugabanywa mububiko bwibitabo bwa peptide, isomero rya cDNA, isomero rya antibody, nububiko bwa protein. Icyiciro cyerekana tekinoroji iroroshye gukora kandi ihendutse gukoresha. Nyamara, ubu buhanga bugabanya itandukaniro rya genetike ya molekuline mu isomero ridashobora kwerekana urutonde rurerure,

Kugeza ubu, tekinoroji yo kwerekana ikoreshwa cyane. Muri byo, hari byinshi byagezweho mu bushakashatsi no guteza imbere inkingo nshya (inkingo zidahenze kandi zikora neza), guteza imbere imiti ya antibody (kwerekana enzyme inhibitori), kwanduza ibimenyetso bya selile (kwerekana epitopi yigana), no gukora ubushakashatsi kuri epitopi ya antigen (gutegura antibodiyite za monoclonal).

Intangiriro kuri T7 Bacteriophage

T7 bacteriophage ni ADN ikubye kabiri, uburebure bwa ADN ni 40kb kandi bupfunyitse muri capsid ifite diameter ya 60nm. Ihuza hagati yumutwe numurizo nuburyo bwimpeta igizwe na kopi nyinshi za gp8. Umutwe nintangiriro ya bacteriophage ya t7 ikora silindrike ihuza gp8 kandi ishobora guhuza umutwe numurizo.

Igishushanyo 1 Igishushanyo cyerekana bagiteri T7.(Reba: Iterambere muri sisitemu ya T7 yerekana (Isubiramo))

Intangiriro kuri M13 Bacteriophage

Indwara ya bacteriophage M13 ni iyitsinda ryibyiciro byiswe hamwe byitwa Ff fage. Ifite uburebure bwa 900 nm n'ubugari bwa 6.5 nm. Irimo genome ya ADN imwe (ssDNA) ifite uburebure bwa 6407 bp igizwe na gen icyenda ikubiyemo poroteyine 11 zitandukanye. Muri byo, poroteyine eshanu ni poroteyine, kandi poroteyine esheshatu zigira uruhare mu kwigana no guteranya icyiciro. Ubwinshi bwa poroteyine za kote ni capsid proteine G8P, igizwe na poroteyine zigera ku 2.700 kandi ikora ibahasha ikikije chromosome.

Igishushanyo cya 2 Igishushanyo mbonera cya bacteriophage M13.(Reba: Shingiro rya Antibody Phage Yerekana Ikoranabuhanga)

Intangiriro kuri Phase Yerekana Isomero rya Antibody

Ubuvumbuzi bwa Antibody bwabaye ingirakamaro mubuvuzi bwa kijyambere. Uburyo butandukanye bwo kuvumbura antibody burahari, ariko fage yerekana isomero rya antibody rikoreshwa cyane mubumenyi bwubuvuzi.

Kuva mu 1990, uburyo butandukanye bwa antibody bwakoreshejwe mukubaka amasomero ya fage, harimo VHs, VHHs, scFvs, diabodies, na antibodies za Fab. ScFv igizwe na VH na VL imiterere yimiterere ni antibodies zumunyururu umwe zikoreshwa mukubaka amasomero ya antibody scFv. scFv irangwa nigice gito cyubuzima hamwe nubudahangarwa buke. Isomero rya Fab Antibody rigizwe na VH, VL, CL, na CH1, rishobora kwerekana vuba antibodi nziza kandi zifitanye isano. Igice gito gishobora guhuza intego antigen-VHH igizwe nububiko bwibitabo bwa nanobody kurubu, VHH ifite ibyiza byuburyo bworoshye, ingano ntoya, gukomera cyane, guhagarara neza, no gutegura byoroshye no kuvuga. Amasomero ya sintetike ya nanobody (Nb) arigaragaza nkuburyo bushimishije bwo gukingira inyamaswa kuri nanobody ihamye, ihuza cyane. Mubisanzwe, ibyo bice bya antibody byahujwe na G3P yicyiciro cya M13, kandi mugukwirakwiza ingirabuzimafatizo nyinshi zigizwe na antibody, isomero rinini ryerekana ububiko bwibitabo bushobora kuvamo antibodi nyinshi zitandukanye.

Icyiciro cyububiko bwibitabo

Igikorwa cyo kubaka isomero rya fage nuburyo bukurikira: primers yihariye yagenewe amplification ya PCR ibona ibicuruzwa, bikaba enzyme ihujwe na vector ya T7 / M13, kandi recombinant phage plasmid yubatswe neza. Fomb plasmid ya recombinant yahinduwe ingirabuzimafatizo za TG1, hanyuma igashyirwa ku gikoresho kirimo antibiyotike ikwiye, kandi umuco wagutse wakozwe nyuma yo gusuzuma transformateur. Nyuma yimico myinshi yumuco, fage yigana ibihe byo kwigana muri bagiteri kandi ikagaragaza neza proteine cyangwa polypeptide. Noneho isomero rya fage risukurwa kugirango rikureho umwanda hamwe nicyiciro kidafunze.

Uturere dutandukanye (VH na VL) dufitanye isano nubwinshi bwa antibody, kandi VH na VL byinjijwe muri vector ya fage hamwe nurutonde rukubiyemo poroteyine ya PIII. Nyuma yo guterana, agace ka fage karashyizwe hamwe hanyuma gahuzwa na N-terminal ya proteine ntoya ya proteine III kugirango ikore igice cya antibody ikora, ibone isomero ririmo antibody ya ADN ikurikirana.

Nyuma yo gukingirwa birangiye, hamenyekanye titer, hanyuma hafatwa amaraso yinyamaswa nyuma yuko titer yujuje ibyangombwa. Lymphocytes yitandukanije n'amaraso, RNA ikuramo, RT-PCR yongerera igice intego, kandi haboneka agace ka V karere. V gene yongerewe imbaraga ikoresheje primer yihariye.

Amasomero karemano yashizweho arimo antibodiyite mbi zo gukingira inyamaswa zishobora kwibasira intego iyo ari yo yose. Antibodiyumu za fage zihuza cyane na antigen zirashobora gukusanywa hakoreshejwe tekinoroji yo gusuzuma ikosora cyangwa ikaranga antigen.

Intego ya antigen ishyizwe mubitwara bikomeye, nk'umwobo wa microplate, cyangwa bigahuzwa n'amasaro ya rukuruzi. Isomero rya antibody ya fage noneho ryongeweho kugirango rihambire kuri antigen. Nyuma yo gutoranya kwinshi, ibyiciro bike cyangwa ibyiciro bidasanzwe byogejwe kandi aberekana antibodi zihariye baragumana.

Gushyira mu bikorwa Icyiciro

* Ubuvumbuzi bwa Antibody bwabaye ingirakamaro mubuvuzi bwa kijyambere. Uburyo butandukanye bwo kuvumbura antibody burahari, ariko tekinoroji yo kwerekana fage ikoreshwa cyane mubumenyi bwubuvuzi. Kuva mu 1990, uburyo butandukanye bwa antibody bwakoreshejwe mukubaka amasomero ya fage, harimo VHs, VHHs, scFvs, diabodies, na antibodies za Fab.
Isomero rya Phage peptide ryemerera kumenya byihuse urukurikirane rwa poroteyine kandi byabaye igikoresho gikomeye cyo gukora iperereza ku mikoranire hagati ya epitopi na reseptor antigen.
* Ibice bya Antibody byahujwe na G3P yo mu cyiciro cya M13, kandi mugukwirakwiza ingirabuzimafatizo nyinshi zigizwe na antibody, isomero rinini ryerekana ububiko bwibitabo bwa antibody bushobora kuvamo antibodi nyinshi zitandukanye.
* T7 yerekana ibyiciro bya T7 ifite ibyiza byinshi, birimo ubworoherane, umutekano muke, umutekano, kubika byoroshye, no gutwara, bityo sisitemu ikoreshwa mugukingira indwara no kuvura.
Sisitemu yo kwerekana fage ya T7 irashobora kumenya antigene zitandukanye, nka antigene yo hejuru ya mikorobe itera indwara na antigene za kanseri.

Icyiciro