Application of semi-industrial monolithic columns for downstream processing of clotting factor IX

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
K BranovićD Josić

Abstract

It has been shown in a previous study that monolithic columns can be used for downstream processing of different concentrates of clotting factor IX [K. Branović et al., J. Chromatogr. A 903 (2000) 21]. This paper demonstrates that such supports are useful tools also at an early stage of the purification process of factor IX from human plasma. Starting with the eluate after solid-phase extraction with DEAE-Sephadex, the use of monolithic columns has allowed much better purification than that achieved with conventional anion-exchange supports. The period of time required for separation is also much reduced. In up-scaling experiments, separations are carried out with 8, 80 and 500 ml columns. A volume of 1830 ml of DEAE-Sephadex eluate, containing a total of 27.6 g of protein and 48500 IU of factor IX is applied to the 500 ml monolithic column. This corresponds to a separation on a pilot scale. The results of this separation after up-scaling are comparable to those obtained with the 8 ml column on a laboratory scale.

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Citations

Aug 11, 2007·Analytical and Bioanalytical Chemistry·N DelmotteC G Huber
Oct 1, 2013·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Daniel A RibeiroLeda R Castilho
Mar 29, 2001·Journal of Chromatography. B, Biomedical Sciences and Applications·D JosicA Jungbauer
Feb 24, 2001·Journal of Chromatography. a·R Hahn, A Jungbauer
Feb 26, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Xindu Geng, Lili Wang
Dec 19, 2006·Journal of Chromatography. a·Djuro Josic, James G Clifton
Sep 4, 2004·Journal of Biochemical and Biophysical Methods·A PodgornikA Strancar
Sep 15, 2017·Electrophoresis·Uroš AndjelkovićMilica Popović
Sep 10, 2004·Journal of Separation Science·Frantisek Svec
Jul 16, 2009·Journal of Separation Science·Michael W H RobertsMichael K Danquah

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