Direct conjugation of bacterial polysaccharides to proteins by squaric acid chemistry

Abstract

Bacterial polysaccharides that contain one amino group can be conjugated using squaric acid chemistry directly to a protein carrier. The conjugation is a two-step process consisting of labeling the polysaccharide with a squarate group and a reaction of the squarate formed with protein. The intermediate squarate derivative and the product glycoconjugate can be easily purified using centrifugal filtration devices. This method is experimentally simple and affords glycoconjugates with predictable carbohydrate–protein ratio (carbohydrate content), high conjugation efficiency, and excellent yield.