Preparation of agarose microspheres with large pore size and high load and purification of antibody in serum based on microfluidic technology

Abstract

The performance of chromatographic media affects the efficiency of purification. In this study, based on a T-type microfluidic droplet generation system, agarose microspheres with large pore size and high load were prepared by adjusting the viscosity of a 4% agarose solution to 70 mPa·s at high temperature. Optical microscopy revealed that the self-made agarose microspheres had a good spherical structure with an average particle size of 82.81 μm and a CV value of 0.0576. Subsequently, the stability and mechanical strength of the self-made agarose microspheres were improved by cross-linking and activation with epichlorohydrin. After the cross-linking and activation, the agarose microspheres were grafted with Protein A to make it have specific adsorption capacity for IgG. The adsorption capacity of the self-made agarose microspheres grafted with Protein A for IgG was 87 mg/g. In the complex serum environment, the agarose microspheres grafted with Protein A still maintained good IgG adsorption capacity and selectivity, demonstrating their potential in practical applications. This study demonstrates the great potential of microfluidic technology in the design and preparation of bio-separation media. By precisely controlling the particle size and pore size of the microspheres, the purification efficiency and selectivity can be significantly improved, providing a new solution for the efficient purification of bio-products.

Graphical abstract