Protein Recovery Using Simulated Moving Bed Technology and Macroporous Chromatographic Support Material

University dissertation from Department of Biotechnology, Lund University

Abstract: Due to its gentle and high resolving power, chromatography is the work horse within protein separation. Still it is an expensive method due to low productivity, diluted product streams, large waste streams and expensive support material. This limits its potential use to areas of high price products. Therefore there is a clear interest to make the technology cheaper to use both for the existing pharmaceutical applications but also to open doors to new areas like processing of effluents from the food industry. The objective of this thesis work was to investigate the potential of simulated moving bed and a new macroporous chromatographic support material in the area of protein separation. The studies presented in this thesis clearly demonstrate that simulated moving bed technology with its inherent coupling flexibility is a very interesting tool for protein isolation, both for individual as well as whole protein isolation. Efforts were made to compare the SMB-technology to single column set-ups in order to investigate whether stated benefits of the SMB could be shown. Perfectly fair comparisons are not easy to make since both processes must be indeed optimized and there should not be differences such as raw material and chromatographic support utilized. Still in the comparisons made it is rather clear that the positive effects of the SMB compared to single column chromatography, can be seen, especially the positive effects regarding buffer saving and product concentration. The other way to improve process economy in chromatography addressed in this thesis was the development of new macroporous chromatographic support material. The macroporous monolithic pAAm-cryogels columns present attractive chromatographic media with unique properties like extremely low flow resistance. The results presented indicate that the important issue of reproducibility for these macroporous monoliths is controlled. Also, the potential of the macroporous monoliths to be used as a capturing column was demonstrated with satisfying results. The studies presented in this thesis are limited to the use of the macroporous material in protein separation. Still there are many possible applications for this new material, especially in the area of chromatographic separation of larger biomolecules and particles like plasmids, viruses and cells. Thus, the full potential of this new support remains to be elucidated.