Fabrication and simulation studies of high-performance anionic sponge alginate beads for lysozyme separation

The large-scale applications of lysozyme in the pharmaceutical industry and food industry require more efficient and cost-effective techniques for its separation. This study present scalable methodology for development of highly porous chromatographic media using an in situ formed network of natural polysaccharide alginate (Alg) through using CaCO3 as solid porogen. The as-prepared porous (Alg) beads demonstrated porous structures with pore diameter 200 nm and 94% porosity. Porous (Alg) beads were functionalized via sodium 3-sulfobenzoate (SS) for lysozyme separation via electrostatic attraction. The optimized sulfonated-alginate beads (SS(8)Alg(0.7)) exhibited excellent lysozyme adsorption capability of 600 mg g-1 within short adsorption time 6 h. More importantly, it shows a dynamic binding efficiency of 500 mg g-1 can be achieved only by the gravity of solution, which matches well with the demands of the high yield and energy conservation in the actual protein purification process. The (SS(8)-Alg(0.7)) beads exhibited high selectivity that enable it directly extract lysozyme from egg white with a relatively large capture capability of 300 mg g-1. Furthermore, the NFM displayed efficient chemical and physical stability even after 5 cycles without significant change in its porous nature or adsorption performance. Significantly, aiming to expand this efficient technique in industrial scale we apply the operating parameters using response surface methodology (RSM) as a modeling tool to optimize the process for further proteins molecules. The premise of this design is that the reusable sulfonate groups are covalently incorporated into the open porous beads structure and can be directly used in reliable separation applications through batch mode or under the driven pressure of gravity.

Automated summary

This study presents a scalable methodology for the efficient separation of lysozyme using highly porous chromatographic media developed through in situ formation of a natural polysaccharide alginate network. The functionalized alginate beads showed excellent lysozyme adsorption capability and high selectivity, making it suitable for industrial scale applications. The optimized beads displayed stability over multiple cycles and the operating parameters were further optimized using response surface methodology for other protein molecules.