Ihuriro rimwe rya selile

Antibody ya Monoclonal (mAb) nk'intangarugero mu miti ya antibody, antibodi zirenga 100 za monoclonal zemerewe kuvura kanseri, indwara zandura, isoko ry’iterambere ry’ibiyobyabwenge, kandi zigaragaza agaciro kihariye ko kuvura. Ikoreshwa rya antibody ya selile imwe ya B, nk'igisekuru gishya cya tekinoroji yo guteza imbere antibody, irashobora gutandukanya neza kandi vuba antibodi mu ngirabuzimafatizo imwe B, ikaba ari intambwe mu buhanga bwo gukora antibody nyuma ya hybridoma na fage yerekanwe. Ikoreshwa rya selile imwe ya selile irashobora gukoreshwa mugukora antibody kurwanya amoko menshi nkinkwavu, imbeba, ingamiya, intama, ninkoko. Tekinoroji ya selile imwe ya B ifite ibyiza byingenzi nkibisobanuro bihanitse, ibikorwa byinshi, hamwe cyane. Ukurikije uburyo bwo gutondekanya ingirabuzimafatizo ya B imwe, Alpha Lifetech ifite ibyiza mugihe cyo gusuzuma no kubona antibodies nziza. Irashobora gutanga igishushanyo cya antigen, synthesis, hamwe no guhindura, ubudahangarwa bwinyamaswa, kugenzura ingirabuzimafatizo imwe ya B, kugenzura ingirabuzimafatizo imwe, kwerekana ibicuruzwa byinshi, kwerekana, no kweza, Guhagarika serivisi tekinike nko kugenzura imikorere kugirango ihuze ibyifuzo bitandukanye byabakiriya.

Inshamake yuburyo bumwe B-Akagari Uburyo bwo Kugenzura

Uburyo bumwe bwo gusuzuma B selile bugabanijwe cyane muburyo bwo kwigunga no kwigunga kwa Antigenselective. Gutandukana bisanzwe birakoreshwa kuburugero hamwe nubushakashatsi bwinshi bwo kwigunga kwa Antigenselective, kandi selile B zonyine ziratandukanye. Nibyoroshye gukora ariko bisaba akazi kenshi, kandi akenshi bikoreshwa nkintambwe yambere yo gutandukana kwihariye kwa antigen.Uburyo bwihariye bwo gutandukana bwa antigen burahuye nibihe ko ibikubiye muri antibodi zihariye nka antibodiyite zirwanya ibibyimba na antibodiyite za autoimmune ari bike. Biragoye cyane gutandukanya selile B yihariye ukoresheje ihame ryikingira rya antigen na antibody ihuza umwimerere. Uburyo bukoreshwa muburyo bwo gutondekanya ibyiza byabwo nibibi bigaragara mumeza.

	Inzira	Ibyiza	Ibibi
Bisanzwe Kwigunga	Micromanipulation	Ibikoresho bike bisabwa Igikorwa cyoroshye	Gukora neza Ubwoko buke bwingirabuzimafatizo
	Lazeri ifata microdissection	Umwanya wo hejuru Igikorwa cyoroshye Biboneka hitamo selile zisa kurugero hanyuma ukureho umwanda	Inzira igoye yo gutegura icyitegererezo Kudashobora gukora
	Fluorescence-ikora ingirabuzimafatizo	Ukuri kwinshi kandi neza Urwego rwo hejuru rwo kwikora Porogaramu nini	Igikorwa kitoroshye Ibikoresho bikenewe cyane
Kwigunga kwa Antigen	Fluorochrome yanditseho antigens ikoresheje Multarameter	Ukuri kwinshi kandi neza Urwego rwo hejuru rwo kwikora Byinshi-byinjira kandi byinshi Ubushobozi bwo gutandukanya B selile mubyiciro bitandukanye	Igikorwa kitoroshye Ibikoresho bikenewe cyane
	Amashanyarazi ya antigencoated	Igikorwa cyoroshye Gusubiramo cyane Irashobora gukungahaza intego B.	Igikorwa kitoroshye Amasaro ya magneti agira ingaruka kumikorere yibinyabuzima ya selile kandi ntabwo afasha umuco nigikorwa nyuma yo kwigunga
	Microengraving	Gukora neza Umuvuduko mwinshi	Ibikoresho bikenewe cyane Gukora neza
	Immunospot array assay kuri chip	Urwego rwo hejuru rwo kwikora Ukuri kwinshi kandi neza	Igikorwa kitoroshye Igiciro kinini

ingirabuzimafatizo B imwe Yerekana Antibody Serivisi

Gukingira inyamaswa

Hitamo antigen ikwiye ku nyamaswa zigeragezwa (nk'imbeba, inkwavu, nibindi) kugirango ukangure ubudahangarwa bw'umubiri, kugirango zishobore gukora antibodi zihariye zirwanya antigen.

Gukingira inyamaswa nicyiciro cyambere cyo kubyara antibody. Gukingira inyamaswa ni ugukoresha uburyo bwo kurwanya ubudahangarwa bw’ibinyabuzima ubwabwo, binyuze mu guhora gukangura antigene, inyamaswa zitanga ubudahangarwa bukomeye bw’umubiri, hanyuma zigasohora antibodi zihariye zirwanya antigene zihariye.

Gukusanya no gutunganyiriza selile B.

Utugingo ngengabuzima B twatandukanijwe n'amaraso ya periferiya, intanga cyangwa lymph node y'inyamaswa zirinda umubiri. Izi selile B zashishikarijwe gukingira antibodiyite zirwanya antigene zihariye.

Ukoresheje uburyo bwumubiri cyangwa bwimiti (nka density gradient centrifugation, gutandukanya amasaro ya magneti, nibindi) gutunganyiriza ingirabuzimafatizo B, kuvanaho selile zanduye, kunoza isuku ya selile B nuburyo bwiza bwo gutandukana nyuma.

Igishushanyo.1 Gukwirakwiza no gutegura antibody ya monoclonal. Inkomoko:Ingirabuzimafatizo imwe ya B hamwe no kubyara inkwavu za Monoclonal Antibodies.)

B gutandukanya selile

Fluorescence-Ikora Utugingo ngengabuzima (FACS), Gutandukanya Akagari ka Magnetique (MACS), Microfluidic Cell Sorter (MCS) cyangwa ubundi buryo bwo kwisuzumisha bwinjira cyane, bufatanije na label yihariye ya antigen, burashobora gukoreshwa mugupima selile B imwe kuva selile ikungahaye kugirango ikore antibodi zigenewe.

Iyo selile imwe B itandukanijwe n’abaturage bavanze B, ingirabuzimafatizo zayo zigomba gukurikiranwa n’ikoranabuhanga rimwe rukurikirana kandi rigasesengurwa kugira ngo haboneke amakuru akurikirana ya ADN y’akarere gakomeye (VH) n’akarere kahinduye urumuri (VL). Ikoranabuhanga rimwe rukurikirana rikubiyemo intambwe eshatu: kwigunga kwingirangingo imwe, kwongera aside nucleic acide selile no gusesengura uko bikurikirana. Ihame ryurwego rukurikirana ni ukongera imbaraga za ADN zose za ADN cyangwa RNA yingirabuzimafatizo imwe kugirango ubone genome yuzuye cyangwa transcriptome ifite ubwuzuzanye bwinshi kugirango bikurikirane byinshi.

Binyuze mu gusuzuma cyane, antibody gene ikurikirana ya selile ibihumbi n'ibihumbi B irashobora kuboneka icyarimwe, byihuta cyane umuvuduko wo kuvumbura antibody. Aya makuru akurikirana arashobora gukoreshwa muburyo bwo kubaka isomero rya antibody no kwerekana.

Igishushanyo.2 Igishushanyo mbonera cyo gufata B selile imwe, kubaka isomero, kwinjizamo ibintu byinshi no gukurikirana.(Inkomoko yerekana: Kuringaniza cyane ingirabuzimafatizo imwe ya B-selile yakira ituma habaho kuvumbura vuba antibodi zitandukanye za antigen-reaction.）

Antibody gene amplification

Akagari kamwe ka B kashizwemo kurekura mRNA. Hanyuma, cDNA ikurikirana yumurongo uremereye uhinduranya akarere (VH) hamwe numurongo uhindura urumuri (VL) byongerewe imbaraga na reaction transcription-polymerase reaction reaction (RT-PCR).

Imvugo ya Antibody no kwezwa

Agace kongerewe imbaraga mukarere gene yinjijwe muri vector plasmid hamwe na gene yo mukarere gahoraho kugirango yubake imvugo ya antibody ya monoclonal. Noneho, uburyo bukwiye bwo kwerekana imvugo (sisitemu yo kwerekana imvugo ya eukaryotic nka selile ya CHO cyangwa sisitemu yo kwerekana imvugo nka Escherichia coli) yatoranijwe kugirango igaragaze, kandi antibody ya monoclonal hamwe nibikorwa byibinyabuzima irabonetse. Poroteyine Chromatografi ya affinity yakoreshwaga mu gukuramo no kweza antibodiyumu zagaragaye mu muco w’akagari no kubona antibodiyite nziza zo mu bwoko bwa recombinant monoclonal.

Kwemeza Antibody

Antibody yemeza umwihariko, ibikorwa hamwe nibikorwa byibinyabuzima byakozwe na ELISA, blot yo muburengerazuba, Flow Analysis, na Immunocoprecipitation kugirango barebe ko antibody ikora neza. Kwemeza Antibody ni intambwe yingenzi mu kwemeza ubwiza no kwizerwa bya antibodies.

Ibisabwa kuri immunogen

. Niba hakenewe kwezwa antigen affinity, hiyongereyeho mg 10 za proteine za recombinant.

. Molekile ntoya isaba isuzumabumenyi hakiri kare.

ingirabuzimafatizo imwe B yo gutondekanya serivisi

Intambwe	Ibirimo	Igihe ntarengwa
Intambwe ya 1: Gukingira inyamaswa no kumenya ELISA	. Muri icyo gihe, 0.1ml ya pre-immunisation na serumu nyuma yo gukingirwa byoherezwa kubakiriya kwipimisha; Niba titer itujuje ibyangombwa, gukingirwa bizakomeza cyangwa umukiriya arasaba guhagarika umushinga, kandi igice cyo gukingira kizishyurwa gusa; (2) Niba titer yujuje ibisabwa, selile PBMC yatandukanijwe namaraso yose.	Ibyumweru 10-12
Intambwe ya 2: Gutandukanya selile imwe	. (2) Ibimenyetso bya Immunoantigen FITC na AF648. . (4) Kumenyekanisha ELISA + ikurikiranye rya clone nziza.	Ibyumweru 2-3
Intambwe ya 3: Imvugo ya Antibody no kwemeza	. (2) Kumenyekanisha ELISA.	Ibyumweru 3-4
Intambwe ya 4: Gutanga	. (2) Buri cyiciro cyerekanwe cyatanze 0.2mg ya antibody. (3) Raporo yubushakashatsi.	Icyumweru 1