A Simple Method for Predicting the Adsorption Performance of Capture Chromatography of Proteins

Abstract

Dynamic binding capacity (DBC) of model proteins in ion-exchange chromatography was analyzed based on a simplified pore diffusion model in order to develop a simple method for designing a capture chromatography process. A dimensionless parameter, F₀＝u₀d_p²/(ZD_m) was derived from the pore diffusion model, where u₀ is the superficial velocity, d_p is the particle diameter, Z is the column bed height, and D_m is the molecular diffusion coefficient. Breakthrough curves of model proteins (lysozyme and immunoglobulin G) were measured for cation exchange chromatography columns of two different particle sizes (d_p＝34 and 90 μm) at various temperatures (8, 13, 18, 23 and 28℃). The efficiency E＝DBC/SBC values calculated from the experimental data were well correlated to F₀ (SBC＝static binding capacity). The E-F₀ curve was found to be useful for designing efficient capture processes of proteins.