Cov khoom

Tshiab ua tau zoo ua kua chromatography assay rau glycosyltransferases raws li derivatization nrog anthranilic acid thiab fluorescence nrhiav tau.

Kev soj ntsuam tau tsim los siv cov cim cim npe ntawm 2-aminobenzoic acid (2AA; anthranilic acid, AA) rau kev ntsuas kev ua ub no ntawm β1-4 galactosyltransferase (GalT-1) thiab α2-6 sialyltransferase (ST-6) los ntawm cov kua ua haujlwm siab. chromatography (HPLC) nrog fluorescence detection (Anumula KR. 2006. Advances in fluorescence derivatization method for high-performance liquid chromatographic analysis of glycoprotein carbohydrates. Anal Biochem. 350:1-23).N-Acetylglucosamine (GlcNAc) thiab N-acetyllactosamine tau siv los ua cov neeg txais thiab uridine diphosphate (UDP)-galactose thiab cytidine monophosphate (CMP)-N-acetylneuraminic acid (NANA) ua tus pub rau GalT-1 thiab ST-6, raws li.Cov khoom lag luam Enzymatic tau sau npe rau hauv qhov chaw nrog AA thiab raug cais tawm ntawm cov substrates ntawm TSKgel Amide 80 kem siv cov xwm txheej ib txwm muaj.Cov enzymes tau txiav txim siab los ntawm thaj chaw ncov los ntawm kev sib piv nrog cov qauv kev sib txuas ua ke ntawm Gal-β1-4GlcNAc thiab NANA-α2-6 Gal-β1-4GlcNAc.Linearity (lub sij hawm thiab enzyme concentration), precision (intra- thiab interassay) thiab reproducibility rau cov kev ntsuam xyuas tau tsim.Cov kev ntsuam xyuas tau pom tias muaj txiaj ntsig zoo hauv kev saib xyuas cov haujlwm enzyme thaum lub sijhawm cais tawm thiab ua kom huv.Cov kev ntsuam xyuas tau muaj kev nkag siab zoo thiab ua tau zoo sib npaug lossis zoo dua li cov kev ntsuas suab thaj hauv cov suab thaj ib txwm muaj.Daim ntawv ntsuam xyuas kuj tseem siv tau los ntsuas kev ua haujlwm ntawm lwm cov kev hloov pauv, yog tias cov neeg txais carbohydrate muaj qhov txo qhov kawg rau kev sau npe.Kev ntsuam xyuas rau glycoprotein txais tau tsim los siv IgG.Ib txoj kev luv luv HPLC profileing tau tsim los rau kev sib cais ntawm IgG glycans (biantennary G0, G1, G2, mono- thiab disialylated), uas pab txhawb kev txiav txim siab ntawm GalT-1 thiab ST-6 cov dej num sai.Tsis tas li ntawd, txoj kev profileing no yuav tsum ua pov thawj muaj txiaj ntsig zoo rau kev saib xyuas cov kev hloov pauv hauv IgG glycans hauv chaw kho mob.