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Transfer of Gradient Chromatographic Methods for Protein Separation to Convective Interaction Media Monolithic Columns

P. Milavec Žmak, H. Podgornik, J. Jančar, A. Podgornik, A. Štrancar

Journal of Chromatography A, 1006 (2003) 195–205

Convective Interaction Media (CIM) columns are monolithic columns optimized for the separation of macromolecules. Some of them operate in the axial mode while others operate in the radial mode depending on the column size. In this work we tested the approach suggested by Yamamoto [Biotechnol. Bioeng., 48 (1995) 444] for transfer of gradient methods between columns of different size. A simplified equation for transfer was derived together with a criterion for its application. Separation was evaluated for a standard protein mixture and peroxidase enzymes present in fermentation broth. Salt and pH gradients were applied. Similar resolutions were obtained for each sample on all columns which demonstrates that the proposed approach can be successfully used for method scale-up on this type of column.

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