Ubwubatsi bw'umubiri (Antibody Engineering) ni iki?

Binyuze mu buhanga bw’uturemangingo tw’umubiri, byashobotse guhindura ingano ya molekile, imiterere ya farumasi, ubushobozi bwo kwirinda indwara, ubushobozi bwo gufatana, imiterere yihariye n’imikorere y’uturemangingo tw’umubiri. Nyuma yo gukora uturemangingo tw’umubiri, gufatana kwatwo mu buryo bwihariye bituma tugira agaciro gakomeye mu gusuzuma no kuvura indwara. Binyuze mu buhanga bw’uturemangingo tw’umubiri, dushobora guhaza ibyifuzo by’imiti n’isuzuma ry’indwara hakiri kare.

Intego y’ubuhanga bw’uturemangingo tw’umubiri ni ugushushanya no gukora imirimo yihariye kandi ihamye uturemangingo tw’umubiri karemano tudashobora kugeraho, bigashyiraho urufatiro rwo gukora uturemangingo tw’umubiri tuvura.

Alpha Lifetech, ifite uburambe bwinshi mu mishinga yayo mu bijyanye no gukora antibody, ishobora gutanga serivisi zihariye zo kurwanya antibody za monoclonal na polyclonal ku moko menshi, ndetse no kubaka no gusuzuma isomero rya antibody za phage display. Alpha Lifetech ishobora guha abakiriya antibody nziza zimeze nk'iz'ibinyabuzima hamwe n'ibicuruzwa bya poroteyine bihuza, ndetse na serivisi zijyanye nayo, kugira ngo habeho antibody nziza, zihariye kandi zihamye. Dukoresheje antibody yuzuye, poroteyine na sisitemu zo kugaragaza phage, dutanga serivisi zijyanye no gukora antibody mu buryo bwo hejuru no mu iherezo, harimo serivisi za tekiniki nko gukora antibody humanization, gutunganya antibody, gukurikirana antibody, no kugenzura antibody.

Icyiciro cy’ibanze cy’ubuhanga bw’uturemangingo tw’umubiri gifitanye isano n’ikoranabuhanga ribiri:

--Ikoranabuhanga rya ADN rihuza

--Ikoranabuhanga rya Hybridoma

Iterambere ryihuse ry’ubuhanga bw’uturemangingo tw’umubiri rifitanye isano n’ikoranabuhanga ritatu ry’ingenzi:

--Ikoranabuhanga ryo gushushanya gene na polymerase chain reaction

--Ikorwa rya poroteyine: Poroteyine zihuza imikorere zikorwa n'uburyo bwo kugaragaza imyitwarire nk'umusemburo, virusi zimeze nk'uduti, n'ibimera

--Igishushanyo mbonera cy'inyubako gifashijwe na mudasobwa

Itsinda rya mbere ry’abasilikare b’umubiri ryahinduwe abantu kugira ngo rikoreshwe mu gukora antibodies za chimeric, aho agace gahinduka k’abasilikare b’imbeba ba monoclonal kahujwe n’agace gahoraho k’uturemangingo twa IgG tw’umuntu. Agace gahuza antigen (CDR) k’umusilikare w’imbeba wa kabiri w’igisekuru cyashyizwe mu bwoko bwa IgG y’umuntu. Uretse agace ka CDR, izindi antibodies zose ni hafi ya antibodies z’umuntu, kandi hashyizweho imbaraga zo kwirinda gutera antibodies z’imbeba z’umuntu (HAMA) mu gihe hakoreshwaga antibodies z’imbeba mu kuvura abantu.

 

Kugira ngo hubakwe isomero ryerekana phage, intambwe ya mbere ni ukubona za jeni zigaragaza ubudahangarwa bw’umubiri, zishobora gukurwa mu tunyangingo twa B tw’inyamaswa zakingiwe (ubwubatsi bw’isomero), zikurwa mu nyamaswa zitakingiwe (ubwubatsi bw’isomero karemano), cyangwa ndetse zigateranywa muri vitro hamwe n’uduce twa jeni za antibody (ubwubatsi bw’isomero rya sintetike). Hanyuma, za jeni zongerwamo na PCR, zigashyirwa muri plasmide, kandi zigashyirwa mu buryo bukwiye bwo kwakira (ubwiyongere bw’umusemburo (ubusanzwe Pichia pastoris), ubwiyongere bw’utunyangingo twa prokaryotic (ubusanzwe E. coli), ubwiyongere bw’utunyangingo tw’inyamabere, ubwiyongere bw’utunyangingo tw’ibimera, n’ubwiyongere bw’utunyangingo twanduye virusi zimeze nk’inkoni). Isanzwe cyane ni sisitemu yo kugaragaza E. coli, ihuza uburyo bwihariye bwo kwandika ubudahangarwa bw’umubiri kuri phage kandi igashyiraho imwe muri poroteyine z’agashishwa ka phage (pIII cyangwa pVIII). Ihuriro rya gene, Kandi rigaragara ku buso bwa bacteriophages. Ishingiro ry’ubu buryo ni ukubaka isomero ryerekana phage, rifite akarusho kuruta amasomero karemano kuko rishobora kugira uburyo bwihariye bwo gufatanya. Nyuma y’ibyo, abasirikare bafite antigen specificity barasuzumwa binyuze mu buryo bwo gutoranya ibinyabuzima, antigens zigamije zirakosorwa, phages zidafite aho zibohewe zigakurwaho inshuro nyinshi, kandi phages zibohewe zigakurwaho kugira ngo zirusheho kwiyongera. Nyuma y’inshuro eshatu cyangwa zirenga, antibody zifite specificity nyinshi na high affinity zirakurwaho.

Igishushanyo cya 3: Kubaka no gusuzuma isomero ry’abarwayi bangiza umubiri

Ikoranabuhanga rya ADN rishobora gukoreshwa mu gukora uduce tw’uturemangingo tw’umubiri. Uturemangingo tw’umubiri twa Fab dushobora guhindurwamo hydrolyze gusa na protease yo mu gifu kugira ngo hakorwe uduce 2 (Fab '), hanyuma tugasya na papain kugira ngo hakorwe uduce twa Fab ku giti cyatwo. Agace ka Fv kagizwe na VH na VL, bifite ubushobozi buke bwo kudahagarara bitewe no kutagira imigozi ya disulfide. Kubwibyo, VH na VL bihuzwa hamwe binyuze muri peptide ngufi ya aside amine 15-20 kugira ngo bikore agace kamwe k’uturemangingo (scFv) gafite uburemere bwa molekile bugera kuri 25KDa.

Inyigo ku miterere y’uturemangingo tw’umubiri muri Camelidae (Camel, LIama, na Alpaca) yagaragaje ko uturemangingo tw’umubiri dufite iminyururu iremereye gusa kandi nta minyururu yoroheje, bityo twitwa uturemangingo tw’umunyururu uremereye (hcAb). Uduce duhinduka tw’uturemangingo tw’umubiri ...

Igishushanyo cya 5: Antibody y'uruhererekane ruremereye na VHH/ Nanobody

Ubushobozi bwo kugaragaza uturemangingo (cell free expression) bukoresha uburyo ADN karemano cyangwa ikoze mu buryo bwa sintetiki kugira ngo bugere ku gukora poroteyine muri vitro, akenshi bukoresha uburyo bwa E. coli expression. Bukora poroteyine vuba kandi burinda umutwaro w’imikorere n’uburozi ku turemangingo iyo bukora poroteyine nyinshi zihuza mu mubiri. Bushobora kandi gukora poroteyine zigoye gukora, nk’izo bigoye guhindura nyuma yo guhindura cyangwa gukora poroteyine zo mu ruhu.