Characterisation of metal-chelate methacrylate monoliths

Journal of Chromatography. a
M PeterkaA Podgornik

Abstract

Monoliths are attractive stationary phases for purification of large biomolecules like proteins because of their flow-unaffected properties. Isolation of histidine containing proteins to high purity can be efficiently performed using metal-chelate interactions within a single chromatographic step. In this work, we investigated properties of commercial metal-chelate methacrylate monoliths-Convective Interaction Media (CIM). Analytical CIM disk monolithic columns and CIM 8 ml monolithic columns were used for purification of tumor necrosis factor-alpha (TNF-alpha) analog LK-801 and green fluorescence protein with 6 histidine tag (GFP-6His). In both cases, purity over 90% was achieved. Dynamic binding capacity at 10% of breakthrough was around 17-18 mg/ml for LK-801 and around 30 mg/ml for GFP-6His. Adsorption isotherm revealed that the maximal capacity is achieved at protein concentration above 60 microg/ml. Dynamic binding capacity and resolution were found to be flow unaffected.

References

Nov 6, 2001·Journal of Biochemical and Biophysical Methods·G S Chaga
Nov 6, 2001·Journal of Biochemical and Biophysical Methods·V Gaberc-Porekar, V Menart
Jun 5, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Fatima M PlievaBo Mattiasson
Mar 24, 2005·Journal of Chromatography. a·Igor MihelicTine Koloini

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Citations

Nov 29, 2012·Analytical and Bioanalytical Chemistry·Erika L PfaunmillerDavid S Hage
Sep 15, 2017·Electrophoresis·Uroš AndjelkovićMilica Popović
Jul 11, 2008·Journal of Separation Science·Oscar G Potter, Emily F Hilder

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