DEVELOPMENT AND OPTIMIZATION OF A SINGLE-STEP CATION CHROMATOGRAPHIC WHEY PROTEIN FRACTIONATION PROCESS: EVALUATION AND COMPARISON OF SCALE-UP STRATEGIES

Abstract This manuscript presents evaluation of various strategies used for the scale-up of chromatographic purification systems. Towards that end, the manuscript provides development and optimization of a single-step whey protein concentrate (WPC) fractionation process at lab scale on a 1.0 × 19 cm column using UNOsphere S. Subsequently, scale-up of this newly developed process and conduction of trials at higher scale is also undertaken by applying various scale-up strategies. For these trials, this work categorized various scale-up factors into constant velocity (CV) and constant bed volume per hour (CBVH) scale-up methods. Scale-up trials in bigger diameter (1.5 cm and 2.5 cm) columns with height to diameter ratio (H/D) =19 and <19 were performed. The effects of changing flow velocity and H/D ratio due to the applied scale-up strategy were evaluated and compared with lab performance in terms of resolution vs. height equivalent to theoretical plate (HETP) relationship. The CBVH method was found to be an efficient method for the scale-up of the developed whey protein resolution system. Also an equation for the prediction of resolution in a scaled column on the basis of a HETP-resolution correlation is presented. It is useful for the prediction of either resolution or bed height in scaled column of WPC or similar biomolecular purification systems.