Urubuga rwo Gutondekanya Seli Imwe ya B

Antibody ya monoclonal (mAb) nk'umuvuzi wa mbere w'imiti irwanya kanseri, antibody zirenga 100 za monoclonal zemejwe kuvura kanseri, indwara zandura, isoko ry'imiti bwite, kandi zigaragaza agaciro kazo kihariye ko kuvura. Ikoranabuhanga rya antibody ya single B cell, nk'igisekuru gishya cy'ikoranabuhanga ryo guteza imbere antibody, rishobora gutandukanya antibody mu buryo bwiza kandi bwihuse uturemangingo twa B, ibi bikaba ari intambwe ikomeye mu ikoranabuhanga ryo gukora antibody nyuma yo kugaragaza hybridoma na phage. Ikoranabuhanga ryo gutondeka uturemangingo twa B rimwe rishobora gukoreshwa mu gukora antibody ku moko menshi nk'inkwavu, imbeba, ingamiya, intama, n'inkoko. Ikoranabuhanga rya single B cell rifite ibyiza bidasanzwe nko kuba ifite umwihariko mwinshi, imikorere myinshi, no gukundana cyane. Hashingiwe ku buryo bwo gutondeka uturemangingo twa Single B, Alpha Lifetech ifite ibyiza mu gihe cyo gusuzuma no kubona antibody nziza. Ishobora gutanga imiterere ya antigen, guhuza, no guhindura, ubudahangarwa bw'inyamaswa, gusuzuma uturemangingo twa B rimwe, gukurikirana uturemangingo, gusuzuma umusaruro mwinshi, no gutunganya, serivisi za tekiniki za One stop nko kugenzura imikorere kugira ngo ihuze n'ibyo abakiriya bakeneye bitandukanye.

Incamake y'Ubuhanga bwo Gupima Selire Imwe ya B

Uburyo bwo gusuzuma aside B imwe bugabanyijemo ahanini uburyo bwo kwitandukanya mu buryo butunguranye n'uburyo bwo kwitandukanya mu buryo butunguranye. Gutandukanya mu buryo butunguranye bikoreshwa ku bipimo bifite icyiciro kinini cya Antigenselective isolation, kandi aside B gusa ni zo zitandukanywa. Biroroshye gukora ariko bisaba akazi kenshi, kandi akenshi bikoreshwa nk'intambwe ya mbere yo gutandukanya aside yihariye. Uburyo bwo gutandukanya aside yihariye bukwiriye iyo ingano y'aside yihariye nka anti-tumor antibodies na anti-immune antibodies ari nke. Biragoye cyane gutandukanya aside B runaka hakoreshejwe ihame ry'ubudahangarwa bw'umubiri rya antigen na antibody specific connecting ku ya mbere. Uburyo bukunze gukoreshwa mu gushyira mu byiciro hamwe n'ibyiza n'ibibi byabwo bigaragazwa mu mbonerahamwe.

	Uburyo	Ibyiza	Imbogamizi
Igitunguranye Kwishyira mu kato	Gukoresha mikoro	Ibisabwa bike ku bikoresho Imikorere yoroshye	Umusaruro muke Ubwoko buke bw'uturemangingo twakuwemo
	Gufata imirasire mito yo gukata hakoreshejwe laser	Ubuhanga bwo hejuru mu myanya Imikorere yoroshye Hitamo uturemangingo gisa n'icyo mu isuzuma hanyuma ukureho imyanda	Uburyo bugoye bwo gutegura icyitegererezo Kudashobora gukora ikoranabuhanga
	Gutondekanya uturemangingo hakoreshejwe fluorescence	Ubuhanga n'imikorere myiza cyane Urwego rwo hejuru rw'imikorere y'ikoranabuhanga Porogaramu yagutse	Igikorwa kigoye Ibikoresho bikenewe cyane
Itandukaniro ry'imisemburo irwanya indwara ya Antigen	Antigens zifite izina rya fluorochrome binyuze mu bipimo byinshi	Ubuhanga n'imikorere myiza cyane Urwego rwo hejuru rw'imikorere y'ikoranabuhanga Ingufu nyinshi n'ibipimo byinshi Ubushobozi bwo gutandukanya uturemangingo twa B mu byiciro bitandukanye	Igikorwa kigoye Ibikoresho bikenewe cyane
	Amasaro ya rukuruzi adafite ibara rya jeni	Imikorere yoroshye Gusubiramo byinshi Ishobora kongera uturemangingo twa B twibasiwe	Igikorwa kigoye Amasaro ya rukuruzi agira ingaruka ku mikorere y'uturemangingo kandi ntafasha mu mikurire no gukora nyuma yo kwitandukanya
	Gushushanya mu buryo bwa mikoro	Umusaruro mwinshi Umuvuduko wo hejuru	Ibikoresho bikenewe cyane Umusaruro muke
	Isuzuma ry'imiterere y'imyunyungugu kuri chip	Urwego rwo hejuru rw'imikorere y'ikoranabuhanga Ubuhanga n'imikorere myiza cyane	Igikorwa kigoye Igiciro kiri hejuru

Uburyo bwo Gusuzuma Ubudahangarwa bw'Agakoko kamwe ka B

Gukingira inyamaswa

Hitamo antigen ikwiye inyamaswa zigeragezwa (nk'imbeba, inkwavu, nibindi) kugira ngo zirusheho gutera imbaraga ubudahangarwa bw'umubiri, kugira ngo zibashe gukora ubudahangarwa bwihariye bwo kurwanya antigen.

Gukingirwa kw'inyamaswa ni cyo cyiciro cya mbere cyo gukora antibody. Gukingirwa kw'inyamaswa ni ugukoresha uburyo bwo gusubiza ubudahangarwa bw'umubiri ubwabwo, binyuze mu gukangura antigens buri gihe, inyamaswa zikora ubudahangarwa bukomeye, hanyuma zigatanga antibody zihariye kuri antigens runaka.

Gukusanya no kongera ubwinshi bw'uturemangingo twa B

Uturemangingo twa B twakuwe mu maraso yo mu mpande, mu ruhu cyangwa mu tunyangingo tw’umubiri tw’inyamaswa zirinda indwara. Utwo tunyangingo twa B twakoreshejwe mu buryo bw’ubudahangarwa bw’umubiri kugira ngo tubone uturemangingo dutera uturemangingo dutera uturemangingo twihariye.

Binyuze mu buryo bw'umubiri cyangwa bwa shimi (nk'uburyo bwo gupima ubucucike bw'ingirabuzimafatizo, gutandukanya uduce duto tw'ingufu, nibindi) kuzuza uturemangingo twa B, gukuraho uturemangingo tw'imyanda, kunoza ubuziranenge bw'uturemangingo twa B no kunoza uburyo bwo gutandukana nyuma yaho.

Igishushanyo cya 1 Gutunganya no gutegura antibody y’urukwavu isanzwe. Inkomoko y’inyandiko:Gutunganya no gukora uturemangingo twa B duto (Single B) tw’urukwavu (Rbbit monoclonal antibodies).)

Gutondekanya utunyangingo twa B

Gusesengura Uturemangingo Dukoresha Fluorescence-Activated Cell Sorting (FACS), Gutandukanya Uturemangingo Dukoresha Magnetic Cell (MACS), Microfluidic Cell Sorter (MCS) cyangwa ubundi buryo bwo gusuzuma hakoreshejwe uburyo bwo hejuru, hamwe n'ibirango byihariye bya antigen, bishobora gukoreshwa mu gusuzuma uturemangingo twa B ku turemangingo twa B twinshi kugira ngo hakorwe uturemangingo twibasiwe.

Iyo aseli imwe ya B yakuwe mu itsinda ry’aseli B rivanze, jeni zayo zigomba gutondekanwa hakoreshejwe ikoranabuhanga ryo gukurikirana aseli imwe no gusesengura kugira ngo haboneke amakuru y’uruhererekane rwa ADN rw’akarere gahinduka k’uruhererekane rurerure (VH) n’akarere gahinduka k’uruhererekane rw’urumuri (VL). Ikoranabuhanga ryo gukurikirana aseli imwe rigizwe ahanini n’intambwe eshatu: gutandukanya aseli imwe, kongera aside nucleic intracellular no gusesengura uruhererekane. Ihame ryo gukurikirana aseli imwe ni uguteza imbere ADN cyangwa RNA ya genome yose y’uturemangingo twihariye kugira ngo haboneke genome yuzuye cyangwa transcriptome ifite ubwinshi bwo gupima uruhererekane rw’ibice byinshi.

Binyuze mu gupima cyane, uruhererekane rw'uturemangingo tw'umubiri tw'uturemangingo twa B ibihumbi rushobora kuboneka icyarimwe, bikongera cyane umuvuduko wo kuvumbura uturemangingo tw'umubiri. Aya makuru y'uruhererekane ashobora gukoreshwa mu kubaka no gusuzuma isomero ry'uturemangingo tw'umubiri.

Igishushanyo cya 2 Ishusho y'ishusho yerekana uburyo bwo gufata uturemangingo twa B kamwe, imiterere y'isomero, gusuzuma no gukurikirana ibintu mu buryo bwo hejuru.(Inkomoko y'aho uherereye: Uburyo bwinshi bwo gukurikirana imiterere y'uturemangingo twa B tw'ingirabuzimafatizo butuma haboneka vuba antibodies zitandukanye zirwanya antigen.)

Kongera imiterere y'uturemangingo tw'umubiri

Akaremangingo kamwe ka B gacibwamo ibice kugira ngo gakure mRNA. Hanyuma, urukurikirane rwa cDNA rw’akarere gahinduka k’uruhererekane rurerure (VH) n’akarere gahinduka k’uruhererekane rw’urumuri (VL) byongerewe imbaraga hakoreshejwe uburyo bwo guhindura uruhererekane rw’uru ...

Ubushobozi bwo gukora no gutunganya ubudahangarwa bw'umubiri

Ingirabuzimafatizo z’akarere zihinduka zashyizwe muri plasmidi y’ingirabuzimafatizo hamwe na gene y’akarere idahinduka kugira ngo hubakwe plasmidi y’ingirabuzimafatizo z ...

Kwemeza ubudahangarwa bw'umubiri

Kwemeza imiterere y’umubiri w’uturemangingo, ubwiyongere n’imikorere yabyo hakoreshejwe ELISA, Western blot, Flow Analysis, na Immunocoprecipitation kugira ngo harebwe ko imikorere y’uturemangingo ihuye n’ibyo twari twiteze. Kwemeza ubwiyongere bw’uturemangingo ni intambwe y’ingenzi mu kwemeza ubuziranenge n’ubudahemuka bw’uturemangingo.

Ibisabwa kugira ngo umuntu ahabwe ubudahangarwa bw'umubiri

(1) Poroteyine y’ibice bivanze: ≥2.5 mg, ubuziranenge bwa poroteyine >85%, uburemere bwa molekile ya poroteyine burenga 10 kDa, ingano ya poroteyine ishongeshwa iri hagati ya 0.5 na 5mg/mL. Niba hakenewe gusukura antigen affinity, hakenewe 10 mg y’inyongera ya poroteyine y’ibice bivanze.

(2) Polypeptides na molekile nto: Peptide yambaye ubusa ≥10 mg, ubuziranenge bwa peptide >90%, ntabwo iri munsi ya 10 AA, ihuza KLH/BSA/OVA cyangwa izindi poroteyine zitwara; Molekile nto zikenera isuzuma ry’imiterere mbere y’igihe.

Inzira y'akazi yo gutondekanya selile imwe ya B

Intambwe	Ibikubiye muri serivisi	Ingengabihe
Intambwe ya 1: Gukingira amatungo no kumenya ELISA	(1) Inyamaswa zakingiwe inshuro 4-5, uhereye ku gipimo cya kane, amaraso yakusanyijwe kugira ngo hamenyekane titer ya antibody ya ELISA (protein antigen ELISA serum titer >10^5; Peptide antigen ELISA serum titer >10^4). Muri icyo gihe, 0.1ml ya serumu mbere yo gukingirwa na nyuma yo gukingirwa yoherezwa ku bakiriya kugira ngo bapimwe; Iyo titer idafite ibyangombwa, urukingo ruzakomeza cyangwa umukiriya agasaba guhagarika umushinga, kandi igice cy’urukingo ni cyo cyonyine kizaba cyishyuzwa; (2) Iyo titer yari yujuje ibisabwa, uturemangingo twa PBMC twatandukanyijwe n'amaraso yose.	Ibyumweru 10-12
Intambwe ya 2: Gutondekanya utunyangingo twa B kamwe	(1) Hakusanyijwe udusimba, hategurwa uburyo bwo gufunga uturemangingo twa B, amaraso yo mu gice cy'imbere arakusanywa, kandi uturemangingo twa PBMC twaratoraguwe. (2) Ibirango bya FITC na AF648 by’ubudahangarwa bw’umubiri. (3) Gutondekanya imiterere y'urumuri rw'amazi (double fluorescence flow sorting) + culture ya single b cell clone. (4) Kumenya ELISA + gukurikirana imiterere y'uturemangingo twanduye.	Ibyumweru 2-3
Intambwe ya 3: Ishyirwa mu bikorwa ry'uturemangingo tw'umubiri no kwemeza	(1) Isesengura rya gene y'uruhererekane rwa 6-10 + kwemeza ikizamini cyo kugaragaza ibimenyetso by'inyamabere. (2) Indangamuntu ya ELISA.	Ibyumweru 3-4
Intambwe ya 4: Gutanga	(1) Utunyangingo tw’umubiri: Utunyangingo 4-6 tw’umubiri (utunyangingo tutaragaragazwa dutangwa hamwe). (2) Buri gice cyagaragaye cyatanze 0.2mg ya antibody. (3) Raporo y'igerageza.	Icyumweru kimwe