Adsorption equilibrium and kinetics of Immunoglobulin G on a mixed-mode adsorbent in batch and packed bed...

It is commonly accepted that efficient protein separation and purification to the desired level of purity is one bottleneck in pharmaceutical industries. MabDirect MM is a new type of mixed mode adsorbent, especially designed to operate in expanded bed adsorption (EBA) mode. In this study, equilibrium and kinetics experiments were carried out for the adsorption of Human Immunoglobulin G (hIgG) protein on this new adsorbent. The effects of ionic strength and pH are assessed. Langmuir isotherms parameters are obtained along with the estimation of the effective pore diffusion coefficient (D_pe) by fitting the batch adsorption kinetics experiments with the pore diffusion model. The maximum adsorption of the IgG protein on the MabDirect MM adsorbent, 149.7±7.1mg·g_dry^-1, was observed from a pH 5.0 buffer solution without salt addition. Adding salt to the buffer solution, and/or increasing pH, decreases the adsorption capacity which is 4.7±0.4mg·g_dry^-1 for pH 7.0 with 0.4M NaCl in solution. Regarding the D_pe estimation, a value of 15.4×10^-6cm²·min^-1 was obtained for a pH 5.0 solution without salt. Increasing the salt concentration and/or the pH value will decrease the effective pore diffusion, the lowest D_pe (0.16×10^-6cm²·min^-1) value being observed for an IgG solution at pH 7.0 with 0.4M NaCl. Fixed bed experiments were conducted with the purpose to validate the equilibrium and kinetic parameters obtained in batch. For a feed concentration of 0.5 g·L^-1 of IgG in pH 5.0 buffer solution with 0.4M NaCl, a dynamic binding capacity at 10% of breakthrough of 5.3mg·g_wet^-1 (15.4mg_IgG·mL_resin^-1) was obtained, representing 62% of the saturation capacity. As far as the authors know, this study is the first one concerning the adsorption of hIgG on this type of mixed mode chromatography adsorbent.