Abstract:Objective Staphylococcus aureus protein A (SPA) is widely used for the purification of monoclonal antibodies due to its high specificity for immunoglobulin G (IgG). However, the high costs of commercial media have limited the further development of SPA. To address this issue, this paper aims to prepare a high-performance affinity chromatography medium based on SPA as the substitute of introduced affinity chromatography media.Method SPA expressed by the recombinant strain was purified via Ni2+ affinity chromatography and then coupled to the Rigose HF matrix to prepare an self-prepared affinity chromatography medium.Result The self-prepared medium had a static saturated adsorption capacity of 115.5 mg/mL (based on the volume of the medium) and a dynamic adsorption capacity of 58.6 mg/mL (based on the volume of the medium). When being used for the separation and purification of monoclonal antibody samples, the self-prepared affinity chromatography medium achieved the purity of over 95%, with a recovery rate of no less than 90%. The medium demonstrated comparable or superior alkali tolerance, reusability, and purification performance for monoclonal antibody samples to commercial media.Conclusion A recombinant strain expressing SPA is successfully constructed, and the optimal preparation conditions of the self-prepared affinity chromatography medium are determined by optimization of the coupling conditions. The self-prepared affinity chromatography medium demonstrates research significance and commercial application value to a certain extent.
