Ibicuruzwa

Amazi mashya akora cyane ya chromatografiya yerekana glycosyltransferase ishingiye kuri derivatisation hamwe na aside anthranilic hamwe na fluorescence.

Ubushakashatsi bwakozwe hifashishijwe imiti idasanzwe ya chimie ya 2-aminobenzoic aside (2AA; aside anthranilic, AA) kugirango bapime ibikorwa byombi β1-4 galactosyltransferase (GalT-1) na α2-6 sialyltransferase (ST-6) n'amazi akora cyane chromatografiya (HPLC) hamwe no kumenya fluorescence (Anumula KR. 2006. Iterambere muburyo bwa fluorescence derivatisation yuburyo bukomeye bwo gusesengura ibintu byamazi ya chromatografique ya karubone ya glycoproteine. Anal Biochem. 350: 1-23).N.Ibicuruzwa bya Enzymatique byashyizweho ikimenyetso hamwe na AA kandi byatandukanijwe na substrate kuri TSKgel Amide 80 inkingi ukoresheje ibihe bisanzwe.Ibice bya Enzyme byagenwe uhereye ahantu hahanamye ugereranije nuburinganire bwakomotse kuri Gal-β1-4GlcNAc na NANA-α2-6 Gal-β1-4GlcNAc.Umurongo (igihe hamwe na enzyme yibanze), ibisobanuro (imbere-hagati) no kubyara kubisubizo byashyizweho.Ubushakashatsi bwagaragaye ko ari ingirakamaro mugukurikirana ibikorwa bya enzyme mugihe cyo kwigunga no kwezwa.Isuzuma ryarakomeye cyane kandi ryakozwe bingana cyangwa byiza kuruta ibipimo gakondo bya radiyo ikora isukari.Imiterere ya assay irashobora kandi gukoreshwa mugupima ibikorwa byizindihererekanyabubasha, mugihe abemera karubone ya hydrata irimo iherezo ryo kugabanya ibimenyetso.Isuzuma ryakira glycoprotein ryakozwe hifashishijwe IgG.Uburyo bugufi bwo gushushanya HPLC bwateguwe mugutandukanya IgG glycans (biantennary G0, G1, G2, mono- na disialylated), byoroheje kugena ibikorwa bya GalT-1 na ST-6 muburyo bwihuse.Byongeye kandi, ubu buryo bwo kwerekana bugomba kwerekana akamaro ko gukurikirana impinduka muri IgG glycans mumiterere yubuvuzi.