Colorimetric determination of N-acetylhexosamine-terminating O-glycosidically linked saccharides in mucins and glycoproteins

摘要: Abstract A sensitive colorimetric assay for detecting mucins and glycoproteins rich in O-glycosidically linked saccharides is reported. The method combines the susceptibility of N-acetylgalactosamine terminating to β-elimination with Morgan-Elson reaction N-acetylhexosamines free reducing ends. All mucin mucin-type but none serum-type tested resulted characteristic color production. gave linear responses range 5 200 μg was also adapted microscale involving use 96-well microtiter plates. microassay which volumes samples reagents are scaled down 2.5-fold particularly useful monitoring mucins, presence other glycoconjugates, large numbers obtained during fractionation procedures. Cesium chloride, cesium bromide, potassium thiocyanate, various detergents do not interfere determination. Guanidine hydrochloride, trifluoroacetate, β-mercaptoethanol decreased by 30 45%; however, interference serious prevent detection their presence. specific gel filtration density gradient centrifugation cystic fibrosis sputum demonstrated.