Distributed nature of equilibrium in affinity chromatography: Mathematical modeling, simulation, and scale-up

A. Chapter I. Studies on scale-up parameters of immunoglobulin separation system using protein A - affinity chromatography. The effects of operational and system parameters on the process scale-up of murine immunoglobulin purification using Protein A - affinity chromatography are investigated. Parameters studied are those related to sample application, elution, ligand concentration on the support matrix, and the change of column size.; B. Chapter II. Distributed nature of equilibrium coefficient in affinity chromatography purification. In affinity chromatography the degree of heterogeneity in the equilibrium between the ligand and product molecules may be one of the most important factors to consider due to the complexity of the forces between two molecules. The importance of understanding the heterogeneous nature of affinity chromatography was emphasized by demonstrating the effect of distributed equilibrium coefficient on the elution concentration profile and the separation performance.; The cases considered were; (1) Non-uniform equilibrium due to the heterogeneity of input product, (2) Distributed equilibrium due to the heterogeneity of ligands on the gel matrices, and (3) Change of the equilibrium during the elution process.; C. Chapter III. Properties of various support matrices and their effect on immuno-affinity purification of protein C. Since the cost of affinity ligands and gel matrices is extremely high any damage to the support matrix during the process increases the total production cost tremendously. Five commercially available support matrices were selected and the parameters of interest for immuno-affinity purification of Protein C were measured and analyzed. The performance of support matrices was evaluated by comparing the productivity calculated from measured parameters.