CARBOHYDRATE AFFINITY PAGE FOR THE STUDY OF CARBOHYDRATE-BINDING PROTEINS

Immobilized neoglycoconjugates covalently cross-linked into a polyacry lamide gel can be used to detect and characterize carbohydrate-binding proteins. The neoglycoconjugates comprise two active groups, sacchari de and allyl, located on a poly(2-hydroxyethylacrylamide) backbone. Th e allyl group cross-links with the polyacrylamide gel matrix, while th e saccharide groups are available for specific protein interactions. T his neoglycoconjugate gel is prepared as a thin layer within the stack ing region of a polyacrylamide gel, and electrophoresis is performed a ccording to native, non-denaturing conditions. Carbohydrate-binding pr oteins, specific for the immobilized neoglycoconjugates, are thus reta rded during electrophoresis, while simultaneously permitting the separ ation of nonbinding proteins according to size and charge. This new ap proach can be used to study carbohydrate-binding proteins in the patho logy of disease or infection.