Serivisi yo guteza imbere ubudahangarwa bw'umubiri bwa monoclonal

Alpha Lifetech Inc.ishobora gutanga uburyo bwo guhanura epitope, gukora peptide, kugaragaza poroteyine, gukora antibody za polyclonal, cyangwa monoclonal ndetse no gutunganya antibody kuva mu mikurire y'uturemangingo, amazi ya ascites cyangwa serum yose.

Alpha Lifetech Inc.ifite uburambe mu guha abakiriya serivisi zitandukanye zihamye zirimo uturemangingo twa hybridoma, imirongo ya genome editing cells, imirongo ya knockdown cells, hamwe n'imirongo ya cells ikora cyane. Tugerageza gutanga serivisi zigezweho ku bakiriya bose kandi serivisi zacu ziremewe kandi zizewe.

Ingamba zo guteza imbere ubudahangarwa bw'umubiri

1. Igishushanyo mbonera n'itegurwa rya antigen
Izi antigen zishobora kuba poroteyine, peptides, cyangwa izindi molekile. Gushushanya ubudahangarwa bw'umubiri bisaba uburyo burambuye buhuza ubumenyi ku miterere ya antigen, ubudahangarwa bw'umubiri, na sisitemu yo gutanga. Alpha Lifetech ifite ikoranabuhanga ryo gushushanya ubudahangarwa bw'umubiri rishobora gushushanya ubudahangarwa bw'umubiri ku bakiriya babukeneye.
Alpha Lifetech itanga poroteyine zivanze ku bakiriya kugira ngo bahitemo. Dufite uburyo butandukanye bwo kugaragaza poroteyine, nka E. coli expression system, yeast expression system, baculovirus-insect expression system, mammalian expression system, nibindi, kugira ngo hakorwe poroteyine zivanze ukeneye mu bushakashatsi.

2. Gukingira inyamaswa
Alpha Lifetech ishobora gutanga ubudahangarwa bw'umubiri ku bakiriya, kandi inyamaswa zirinda indwara nk'imbeba, inkwavu, intama, alpacas, guinea pigs, hamsters, n'izindi zirahari kugira ngo uhitemo.

3. Uburyo bwo gukora antibody imwe mu mubiri

●Umusaruro wa Mabs ukoresheje ikoranabuhanga rya Hybridom
Hybridoma ni selile ivanze ikorwa no guhuzwa kw'ubwoko bubiri bw'uturemangingo, ari bwo umurongo w'uturemangingo twa myeloma y'imbeba n'uturemangingo twa plasma (lymphocyte B), aho selile ya heterozygous ishobora gukora antibody zikomeza kurwanya antigen ishimishije muri vitro. Gukora umurongo w'uturemangingo twa hybridoma bigizwe n'ibice bitatu: imiterere ya antigen (haptens, molecules nto, na peptides), gukingira inyamaswa, guhuza uturemangingo, no gusuzuma neza clone. Dukoresheje uburyo bwihariye bwo gukingira, tubona ubushobozi budasanzwe bwo guhuza uturemangingo (hybridomas 1-10 kuri buri turemangingo igihumbi twa B) n'uturemangingo twinshi, bikongera igipimo cyacu cyo gutsinda mu bizamini bikurikira birimo gusuzuma biochemical, isuzuma rya selile muri vitro, n'inyigo ku nyamaswa.

●Umusaruro wa Mabs ukoresheje ikoranabuhanga ryo kwerekana phage
Ishusho ya Phage itanga ubundi buryo bwo gukora antibodies za monoclonal. B-lymphocytes zikurwa mu maraso y'inyamaswa hanyuma mRNA yazo igakurwamo. Binyuze muri PCR, iyi mRNA ihinduka cDNA, yongera ibice byose bya VH na VL. Ibi bice bishyirwa mu cyuma gihindura imiterere, ubusanzwe nk'uduce duhinduka tw'uruhererekane rumwe (scFv), hamwe na poroteyine ya PIII ya bacteriophage. Nyuma yaho, iyi shusho ikoreshwa mu kwanduza E. coli, bigatuma hakorwa isomero ririmo uturemangingo tugera kuri 10^10 binyuze mu gutera inshinge hifashishijwe phage ifasha. E. coli ishobora gusohora bacteriophage ifite ibice bya VH na VL nk'igice cy'umubiri wayo. Nyuma y'ibi, ibice byihariye bya VH na VL byibanda kuri antigen ishimishije bishobora gutoranywa no gukoreshwa mu kongera gukurura E. coli hamwe na bacteriophage. Uturemangingo tugizwe na plasmid turakurwamo kandi tugashyirwa mu byiciro.

4. Gusuzuma imikoranire

●Guhuza no Guhitamo Uturemangingo
Hanyuma uturemangingo twa B twasaruwe duhuzwa n’uturemangingo twa myeloma, ari two turemangingo twa kanseri dushobora kwiyongera mu buryo budashira mu mikurire. Ubusanzwe iyo mikurire ikorwa hakoreshejwe uburyo nka polyethylene glycol (PEG) fusion.
Nyuma yo kwivanga, uturemangingo twa hybridoma duhingwa mu buryo bwo guhitamo butuma hybridomas gusa zikomeza kubaho. Ubusanzwe ubwo buryo bukubiyemo aminopterin, ibuza gukura kwa myeloma zitavanze.

●Gusuzuma no Gufunga
Uturemangingo twa hybridoma dusohoka turasuzumwa kugira ngo harebwe niba hari uturemangingo twihariye kuri antigen. Ibi bikorwa hakoreshejwe uburyo nka enzyme-linked immunosorbent assay (ELISA), Western blot, immunoprecipitation, na flow cytometry.

Iyo uturemangingo twa hybridoma dukora antibody tumenyekanye, dushyirwa mu bwoko bwa clone kugira ngo habeho uturemangingo tumwe. Ibi bituma antibody ikorwa na buri turemangingo twa hybridoma iba imwe.

5. Igenzura ry'imikorere y'ubudahangarwa bw'umubiri
Alpha Lifetech itanga uburyo bwo kugenzura imikorere y’uturemangingo tw’umubiri, nko gusuzuma ibidukikije bya Western, ubutabire bw’umubiri, cyangwa ibizamini by’imikorere, kugira ngo hamenyekane imiterere y’uturemangingo tw’umubiri tw’umubiri w’umuntu umwe kandi hakemezwe imiterere y’uturemangingo tw’umubiri, imiterere y’uturemangingo, n’imikorere yatwo.

6. Guteza imbere ubudahangarwa bw'umubiri
Gusukura antibody zivuye mu musaruro w’umubiri hakoreshejwe uburyo nka poroteyine A cyangwa poroteyine G affinity chromatography, na Alpha Lifetech ishobora no gutanga uburyo bwo guhindura antibody kugira ngo inoze antibody affinity.

7. Gukora ubudahangarwa bw'umubiri
Alpha Lifetech ishobora gusuzuma antibodies zishobora gukoreshwa mu gusuzuma umusaruro mwinshi no gukora ku bwinshi.

Ibibazo Bikunze KubazwaIbibazo Bikunze Kubazwa

IBIBAZO BIKUBIYEMO KU BIBAZO BYA MBERE KU MITERERE Y'AMAVUKO Y'UMUBIRI W'AMAVUKO

1

Ni gute wahitamo fusion hybridoma ikwiye?

Mu gihe cyo guhuza, amasahani 10-15 ya microtiter afite ibiziba 96 aterwamo uruvange rwa fusion hybrid. Buri sahani ihabwa HAT-IMDM selection media kandi ikabikwa mu cyuma gishyushya karubonike gaze kuri dogere selisiyusi 37. Nyuma y'iminsi 9-14, supernatants za hybrid zipimwa niba hari antibody runaka ikenewe.
2

Ni gute wakongera imikorere myiza mu guhitamo?

Guhuza uturemangingo twa plasma n’utundi duto twa myeloma ntabwo bigira ingaruka nziza 100%. Nubwo haba hari ibintu byiza kandi bigatera imbaraga nyinshi, guhuza uturemangingo biracyakora uruvange rw’uturemangingo tudahuye n’utundi duto tugomba gutandukana. Kugira ngo hongerwe imikorere myiza y’uburyo bwo gutoranya, uturemangingo twa myeloma dukoreshwa mu guhuza uturemangingo tutagira HGPRT, enzyme y’ingenzi mu nzira yo kurokora nucleotide. Hanyuma uruvange rwashyizwe mu buryo bwa HAT, aho uturemangingo dufite enzyme ya HGPRT, hybridomas zarazwe HGPRT n’uturemangingo twa plasma, ari two twashoboraga kubaho.
3

Ni gute wasuzuma imirongo y'uturemangingo twa hybridoma kugira ngo urebe niba antibody ikora?

Isuzuma ry'ubwoko bw'ibimera bivanze ni intambwe y'ingenzi. Ubusanzwe, gusuzuma ibimera bivanze, ibimera bivanze, n'ibimera bivanze bikorerwa hifashishijwe isuzuma rya enzyme-linked immunosorbent assay (ELISA), isuzuma rya Western blot, n'isuzuma rya fluorescence activated cell sorting (FACS). Tuzahitamo gukora isuzuma ryose rya ibimera bivanze muri laboratwari yacu.
4

Tanga amabwiriza arambuye y'uburyo bwo gusuzuma ibikorwa by'abarwanyi b'umubiri?

Indwara za Hybridom supernates zigomba gupimwa zishobora kuva ku bipimo 500 kugeza ku 1.440 kandi zizaba ziteguye gupimwa ku munsi wa 9-14. Dushobora kugaragara mu gihe cy'iminsi itatu kugeza kuri itanu cyangwa twese tukaba twiteguye umunsi umwe, bityo ni ngombwa ko laboratwari y'abakiriya iba yiteguye ibyumweru mbere y'uko isuzuma ryiyongera rikorwa.
5

Kuki hybridoma nziza zigomba kwibasirwa n'uturemangingo?

Nyuma yuko Wells zagaragaye mu gihe cyo gusuzuma ELISA bwa mbere, hybridomas zoherejwe mu bwinshi, ni ukuvuga plaque 24-visit, mu rwego rwo kwitegura inzira yo kuzigabanya. Gushyira hybridomas mu buryo bwo kuzigabanya akenshi bikorwa hakoreshejwe uburyo bwo kuzigabanya kugira ngo habeho kuzitandukanya monoclones zihamye. Ubu buryo burimo kuzigabanya no kuzikwirakwiza mu plaque 96-visit kugira ngo habeho monoclonality (akaremangingo kamwe kuri buri plaque). Nibura hagomba gukorwa uburyo bubiri bwo kuzigabanya kugira ngo hagabanuke ibyago byo kurwara hybridomas zivanze.
6

Ni izihe nyungu zo gukoresha ikoranabuhanga rya hybridoma mu kuvumbura antibody?

Uturemangingo tw’ubwoko bwa hybrid twashimiwe ubushobozi bwatwo bwo gukora antibodies zifite ubwiyongere, ubusugire, kandi zihariye mu buryo buhendutse cyane.