KIT FOR SACCHARIDE-RELATED ENZYME ACTIVITY DETERMINATION AND ACTIVITY DETERMINATION USING THE SAME

摘要

PURPOSE:To accomplish the title determination simply, accurately and in high sensitivity by incorporating the reduced terminal of a saccharide with a saccharide PA derivative with 2-pyridylamino group directly bound thereto. CONSTITUTION:About 10nmol of a saccharide PA derivative of formula I (G is galactose; GN is N-acetylglucosamine; Glc is a compound of formula II) is dissolved in ca.1ml of a 0.1M of sodium cacodylate buffer solution (pH ca.6.8) to obtain the objective kit for determining the activity of saccharide-related enzyme (e.g. endo-beta-galactosidase). Second, (1) 0.1-10mM of a sugar nucleotide (e.g. cytidine 5'-phosphate-N-acetylneuraminic acid) as donor and (2) a specimen containing a saccharide-related enzyme (e.g. blood, urine) are added to this kit followed by reaction at 10-50 deg.C and pH4-9 into a reaction product. Third, this reaction product is adsorbed and separated using e.g. an ion exchange technique followed by eluation. Thence, the resulting component is detected using e.g. a fluorescence detection technique, thus determining saccharide-related enzyme activity.