Preparation and characterization of highly porous cellulose-agarose composite chromatographic microspheres for enhanced selective separation of histidine-rich proteins

•The prepared microspheres possess a homogeneous covalent bonding network.•The prepared microspheres show high specific surface area (183.81 m2/g).•The Cu2+ modified microspheres show high performance to histidine-rich proteins. In this work, the porous cellulose-agarose microspheres with high specific surface area and enhanced mechanical strength are prepared by a novel chemical crosslinking method. The crosslinking reaction homogeneously proceeds between polysaccharides, and the covalent bonding network is generated to replace the inherent hydrogen bonding network of cellulose. The prepared microspheres exhibit low crystallinity of 12.45%, which means high content of amorphous regions. The micro-meso-macroporous structure of microspheres in morphology is conducive to high permeability and adsorption capacity, and the microspheres possess high specific surface area of 183.81 m2/g. The affinity chromatographic microspheres are prepared by immobilizing Cu2+, which exhibits high adsorption capacity of 197.65 mg/g for bovine hemoglobin (BHb), fast adsorption rate wihin 40 minutes, well-selectivity, and excellent recyclability in ten cycles. We expect that this work to provide an outstanding candidate for the high performance of biomacromolecular purification.