Sequence analysis of saccharide material

Abstract

A method of analyzing and sequencing saccharide material composed of saccharide chains. The saccharide chains are end referenced, e.g., by labeling or tagging at their reducing ends, and the saccharide material is subjected to a controlled partial depolymerisation using a selection scission reagent, for example, low pH nitrous acid, which cleaves internal glycosidic linkages in accordance with known linkage specificity so as to produce a mixed set of saccharide chain fragments having different lengths ranging throughout the full spectrum of possible lengths for the particular glycosidic linkage specificity of the selective scission reagent employed. Samples of the mixed set of saccharide chain fragments are then treated with selected exoenzymes including exoglycosidases that cleave only particular glycosidic linkages at the non-reducing end of saccharide chains. These exoenzymes are applied to the samples either singly or in combination in accordance with a predetermined strategy. The treated samples are then analyzed, to detect the chain fragments present which have a reducing end derived from the reducing end of the corresponding chain in the original saccharide material and the monosaccharide sequence in the saccharide starting material is then deduced.