Serivisi ishinzwe iterambere rya Monoclonal

Yamazaki Inc.Irashobora gutanga ubuhanuzi bwa epitope, synthesis ya peptide, imvugo ya poroteyine, umusaruro wa antibody ya polyclone, cyangwa monoclonal kimwe no kweza antibody mu muco w’akagari, itera amazi cyangwa serumu yose.

Yamazaki Inc.ni inararibonye mugutanga abakiriya serivisi zinyuranye zihamye zirimo selile ya Hybridoma, genome yo guhindura imirongo, imirongo ya knockdown, hamwe nimirongo ikabije. Turagerageza gutanga serivise yihariye ya serivise kumurongo kuri buri mukiriya kandi serivisi zacu zaragaragaye kandi zizewe.

Ingamba zo Gutezimbere Antibody

1.Igishushanyo mbonera cya antigen
Izi antigene zirashobora kuba poroteyine, peptide, cyangwa izindi molekile. Gutegura immunogène bisaba uburyo bwuzuye buhuza ubumenyi bwimiterere ya antigen, immunologiya, hamwe na sisitemu yo gutanga. Alpha Lifetech ifite tekinoroji ya antigen ishobora gukora immunogène kubakiriya bakeneye.
Alpha Lifetech itanga ibicuruzwa bya poroteyine recombinant kubakiriya bahitamo. Dufite uburyo butandukanye bwo kwerekana poroteyine, nka sisitemu yo kwerekana imvugo ya E. coli, sisitemu yo kwerekana imisemburo, sisitemu yo kwerekana udukoko twa baculovirus, sisitemu yo kwerekana inyamaswa z’inyamabere, n'ibindi, kugirango tubyare proteine za recombinant ukeneye ubushakashatsi.

2. Gukingira inyamaswa
Alpha Lifetech irashobora gutanga ubudahangarwa kubakiriya, kandi inyamaswa zirinda umubiri nkimbeba, inkwavu, intama, alpaka, ingurube, ingurube, nibindi birahari kugirango uhitemo.

3.Uburyo bwo gukora antibody ya monoclonal

●Mabs Umusaruro ukoresheje tekinoroji ya Hybridoma
Hybridoma ni selile ivangwa noguhuza ubwoko bubiri bwingirabuzimafatizo, arizo imbeba myeloma selile na plasma selile (lymphocyte B), aho selile ya heterozygous ishobora gukora antibodies zihoraho zirwanya antigen yinyungu za vitro. Umusaruro wumurongo wa Hybridoma ugizwe nibice bitatu: igishushanyo cya antigen (haptens, molekile nto, na peptide), gukingira inyamaswa, guhuza ingirabuzimafatizo, no gusuzuma clone nziza. Twifashishije uburyo budasanzwe bwo gukingira, tubona imbaraga zidasanzwe za selile-fusion (1-10 ya Hybridoma kuri selile igihumbi B) hamwe na titerite ya antibody nyinshi, bigatuma twatsindira cyane mubizamini byakorewe isesengura ririmo ibizamini bya biohimiki, ubushakashatsi bwakozwe na vitro-selile, hamwe nubushakashatsi bwinyamaswa.

●Mabs Umusaruro ukoresheje tekinoroji yo kwerekana
Icyiciro cyerekana ubundi buryo bwo kubyara antibodiyite za monoclonal. B-lymphocytes itandukanijwe namaraso yinyamaswa hanyuma mRNA ikuramo. Binyuze muri PCR, iyi mRNA ihindurwa muri cDNA, yongerera ibice byose VH na VL. Ibi bice noneho byegeranijwe mubice, mubisanzwe nkibice bitandukanye byumunyururu (scFv), hamwe na poroteyine ya PIII ya bagiteri. Ibikurikiraho, iyi nyubako ikoreshwa mu kwanduza E. coli, bituma hashyirwaho isomero ririmo selile zigera kuri 10 ^ 10 binyuze mu gutera inshinge zifasha. E. coli noneho irashobora gusohora bacteriophage ibitse ibice bya VH na VL nkigice cyayo. Nyuma yibi, ibice byihariye bya VH na VL byibasiye antigen yinyungu birashobora gutoranywa no gukoreshwa muguhuza E. coli na bagiteri. Ingirabuzimafatizo zirimo plasmid noneho zitandukanijwe kandi zikurikiranye.

4.Gusuzuma

●Guhuza Akagari no Guhitamo
Utugingo ngengabuzima B twasaruwe noneho duhujwe na selile myeloma, ari selile kanseri ishobora kwiyongera ubuziraherezo mu muco. Ihuriro risanzwe rigerwaho hifashishijwe tekinike nka polyethylene glycol (PEG).
Nyuma yo guhuza, ingirabuzimafatizo za Hybridoma zifite umuco muburyo bwatoranijwe butuma imvange zibaho gusa. Ubusanzwe uburyo burimo aminopterine, irinda imikurire ya myeloma idakoreshwa.

●Kugaragaza no Gufunga
Ingirabuzimafatizo za Hybridoma zapimwe kugirango zivemo antibodies zihariye antigen igenewe. Ibi mubisanzwe bikorwa hakoreshejwe tekinoroji nka enzyme ihujwe na immunosorbent assay (ELISA), blot yo muburengerazuba, immunoprecipitation, hamwe na cytometrike.

Ingirabuzimafatizo za Hybridoma zimaze gukora antibody zimaze kumenyekana, zirakoronijwe kugirango zitange umubare w'ingirabuzimafatizo zimwe. Ibi byemeza ko antibody yakozwe na buri selile ya hybridoma isa.

5.Kugenzura imikorere
Alpha Lifetech itanga antibody ikora neza, nka Western blotting, immunohistochemie, cyangwa assay imikorere, kugirango iranga antibodiyite ya monoclonal no kwemeza antibody yihariye, isano, n'imikorere.

6.Gutezimbere umuntu
Isuku rya antibodies ziva mumico ndengakamere ukoresheje tekinoroji nka protein A cyangwa protein G affinity chromatography, na Alpha Lifetech irashobora kandi gutanga uburyo bwo guhindura antibody kugirango tunonosore antibody.

7.Umusaruro udasanzwe
Alpha Lifetech irashobora gusuzuma antibodiyite zabakandida kubikorwa byinshi-byerekana ibicuruzwa hamwe n’umusaruro munini.

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Ibibazo byiterambere rya monoclonal antibody

1

Nigute ushobora guhitamo Hybridoma ibereye?

Mugihe cyo guhuza, 10-15 96-isahani ya microtiter isa neza hamwe nuruvange rwa fusion. Buri sahani igaburirwa itangazamakuru ryatoranijwe rya HAT-IMDM kandi rikabikwa muri karuboni ya dioxyde de carbone kuri dogere selisiyusi 37. Iminsi 9 - 14 nyuma yo guhuza, supernatants ya Hybrid irageragezwa kugirango habeho antibody yihariye yinyungu.
2

Nigute ushobora kunoza imikorere yo guhitamo?

Guhuza selile ya plasma hamwe na myeloma bagenzi babo ntabwo ikora neza 100%. Ndetse no mubihe byiza hamwe no gukangura cyane, guhuza selile biracyatanga uruvange rwingirabuzimafatizo zidafite akamaro kandi zigomba gutandukana. Kunoza imikorere yuburyo bwo gutoranya, selile ya myeloma ikoreshwa muguhuza ibura HGPRT, enzyme yingenzi mumihanda ya salvage nucleotide. Uruvange rwahise rutezwa imbere muburyo bwa HAT, aho selile gusa zifite enzyme ya HGPRT, hybridoma yarazwe HGPRT kuva selile plasma, byari byiza.
3

Nigute ushobora kwerekana imirongo ya selile ya Hybridoma kubikorwa bya antibody?

Isuzuma ryubwoko bwimvange nintambwe yingenzi. Ubusanzwe, Hybridoma, clone, na subclone isuzuma ndengakamere itangwa binyuze mumisemburo ifitanye isano na immunosorbent assay (ELISA), blot assay yo muburengerazuba, hamwe na fluorescence ikora selile (FACS). Tuzahitamo gukora ibizamini byose byindengakamere muri laboratoire yacu.
4

Tanga amabwiriza arambuye yuburyo bwo gukora antibody ibikorwa byo gusuzuma?

Indwara ya Hybridoma igomba gupimwa irashobora kuva kuri 500 kugeza kuri 1,440 kandi izaba yiteguye kwipimisha muminsi 9- 14. Turashobora kuvuka mugihe cyiminsi itatu cyangwa itanu cyangwa twese dushobora kuba twiteguye kumunsi umwe, kubwibyo rero ni ngombwa ko laboratoire yabakiriya itegurwa ibyumweru mbere hamwe nogukora icyiciro cya kabiri cyo gusuzuma.
5

Kuki Hybridoma nziza ikeneye guhagarikwa?

Nyuma yuko Wells nziza zamenyekanye mugihe cyambere cyo gusuzuma ELISA, hybridoma yimuriwe mubunini bunini, ni ukuvuga amasahani 24-meza, mugutegura uburyo bwo gutabaza. Subcloning ya hybridoma isanzwe ikorwa nuburyo buke bwo guhinduranya kugirango habeho kwigunga kwa monoclone ihamye. Tekinike ikubiyemo kugabanya imico ya hybridoma no kuyisasa mu byapa 96-byiza kugirango ugere kuri monoclonality (selile imwe kuri buri riba). Nibura hagomba gukorwa ibice bibiri bigabanya kugabanya ibyago byo kwandura abaturage bavanze.
6

Ni izihe nyungu zo gukoresha tekinoroji ya hybridoma mu kuvumbura antibody?

Imirongo ya selile ya Hybrid yashimiwe kubushobozi ifite bwo gukora antibodies zifitanye isano ryinshi, ituze, kandi yihariye muburyo buhenze cyane.