Alaabta

Qiimaynta chromatography dareere wax-qabad cusub oo cusub oo loogu talagalay glycosyltransferases oo ku salaysan kala saarista asiidhka anthranilic iyo ogaanshaha fluorescence.

Qiimaynta waxaa la sameeyay iyadoo la adeegsanayo kiimikada calaamadaynta gaarka ah ee 2-aminobenzoic acid (2AA; anthranilic acid, AA) si loo cabbiro hawlaha labadaba β1-4 galactosyltransferase (GalT-1) iyo α2-6 sialyltransferase (ST-6) ee dareeraha waxqabadka sare leh. chromatography (HPLC) oo leh ogaanshaha fluorescence (Anumula KR. 2006. Horumarka hababka soosaarida fluorescence ee falanqaynta chromatographic dareeraha waxqabadka sare leh ee karbohaydraytyada glycoprotein. Dabada Biochem. 350: 1-23).N-Acetylglucosamine (GlcNAc) iyo N-acetyllactosamine ayaa loo isticmaalay sidii aqbalayaasha iyo uridine diphosphate (UDP) -galactose iyo cytidine monophosphate (CMP) -N-acetylneuraminic acid (NANA) sida deeq-bixiyeyaasha GalT-1 iyo ST-6, siday u kala horreeyaan.Alaabooyinka Enzymatic waxaa lagu calaamadeeyay goobta AA waxaana laga soocay substrates-ka tiirka TSKgel Amide 80 iyadoo la adeegsanayo xaaladaha caadiga ah.Unugyada enzyme waxaa lagu go'aamiyey meelaha ugu sarreeya marka la barbar dhigo heerarka isku dhafan ee Gal-β1-4GlcNAc iyo NANA-α2-6 Gal-β1-4GlcNAc.Linearity (waqtiga iyo feejignaanta enzyme), saxnaanta (intra-iyo interassay) iyo dib u soo saarista imtixaanada ayaa la aasaasay.Tijaabooyinku waxay ogaadeen inay faa'iido u leeyihiin la socodka hawlaha enzymes inta lagu jiro go'doominta iyo nadiifinta.Tijaabooyinku waxay ahaayeen kuwo aad xasaasi u ah oo la sameeyay si la mid ah ama ka fiican marka loo eego cabbirada sonkorta shucaaca ee caadiga ah.Qaabka qiimaynta waxa kale oo loo isticmaali karaa in lagu cabbiro dhaqdhaqaaqa xawilaadaha kale, waase haddii aqbaleyaasha karbohaydraytyadu ay ku jiraan dhammayn yaraynta calaamadaynta.Qiimaynta aqbalayaasha glycoprotein ayaa la sameeyay iyadoo la adeegsanayo IgG.Hab gaaban oo HPLC ah ayaa loo sameeyay kala soocida IgG glycans (biantennary G0, G1, G2, mono-iyo diialylated), kaas oo fududeeyay go'aaminta hawlaha GalT-1 iyo ST-6 si degdeg ah.Intaa waxaa dheer, habkan sifayntu waa inuu noqdaa mid waxtar u leh la socodka isbeddelada IgG glycans ee goobaha caafimaadka.