Prediction of the performance of preparative affinity chromatography

This paper describes a theoretical approach to the prediction of the performance of preparative affinity separations for biological macromolecules in packed columns. The approach, which is applicable to conventional low-pressure packed column methods as well as high-performance liquid chromatography techniques, requires knowledge of certain parameters that describe the interactions between adsorbent and adsorbate during the affinity separation procedure. We have measured the parameters appropriate to the adsorption stages of affinity systems involving immobilised Cibacron Blue and immobilised monoclonal antibodies against beta-galactosidase. The theoretical predictions appear to agree well with the experimental performance of batch and packed column affinity systems. The influence of the factors that govern the performance of the adsorption stage of the separation procedure is explained in detail, and the possible advantages of using HPLC techniques in macropreparative affinity chromatography are discussed.