Double-emulsion templated macroporous cellulose microspheres as a high-performance chromatographic media for protein separation

Cellulose microspheres are commonly chromatographic media yet seriously limited in biomacromolecules separation and purification due to the slow mass transfer kinetics resulting from their narrow nanopores. Herein, a macroporous cellulose microsphere (MCM) with enhanced mass transfer ability has been successfully developed by an oil-in-water-in-oil (O 1 /W/O 2 ) double emulsion strategy. The evolution profile of the double emulsion was tracked and achieved the optimization of interconnected macroporous structure. The macroporous structure not only provide fast mass transfer pathways for proteins but also increase the accessibility of meso/micropores. Benefitting from the macropores, the obtained diethylaminoethyl-modified MCM (DEAE-MCM) exhibits high permeability (3.81 × 10 –13  m 2 ), and fast adsorption rate (reaching equilibrium within 40 min) and high adsorption capacity (334.21 mg/g) for bovine serum albumin, far superior to commercially DEAE Sepharose Fast Flow. More importantly, under the high flow rate, DEAE-MCM remains a high dynamic adsorption capacity, promising it for fast protein chromatography.