Carbohydrate-Coupled Sepharose 6B Matrix Preparation

Carbohydrate coupled Sepharose 6B matrix was prepared according to our previous study (24 (link)). In brief, epoxy-activated Sepharose 6B matrix (GE Healthcare, Sweden) was washed extensively with distilled water, and then mixed with four kinds of carbohydrates, including d-lactose, N-acetyl-d-glucosamine (d-GlcNAc), d-mannose, and l-fucose (Sigma-Aldrich, Buchs, Switzerland), at a final concentration of 200 µM in distilled water (pH 13.0). The mixtures were incubated at 30°C with gentle shaking for 16 h, and 1 M ethanolamine (pH 8.0) was used to block the remaining active groups after TBS washing. The carbohydrate coupled Sepharose 6B matrix was then washed extensively with TBS, and packed into Teflon columns with 100 mm long and 10 mm inner diameter, respectively.