Author/Editor     Milavec-Žmak, P; Podgornik, H; Jančar, J; Podgornik, A; Štrancar, A
Title     Transfer of gradient chromatographic methods for protein separation to convective interaction media monolithic columns
Type     članek
Source     J Chromatogr A
Vol. and No.     Letnik 1006
Publication year     2003
Volume     str. 195-205
Language     eng
Abstract     Convective Interaction Media (CIM) columns are monolithic columns optimized for the separation of macromolecules. Sotne 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.
Descriptors     CHROMATOGRAPHY, HIGH PRESSURE LIQUID
PROTEINS
MACROMOLECULAR SYSTEMS