Elution of antibodies from a Protein-A column by aqueous arginine solutions

Protein Expression and Purification
Tsutomu ArakawaDaisuke Ejima

Abstract

Acidic pH is commonly used to elute antibodies from Protein-A affinity column, although low pH may result in aggregation of the proteins. As an alternative, here arginine was tested as an eluent and compared with a more conventional eluent of citrate. Using purified monoclonal antibodies, recovery of antibodies with 0.1M citrate, pH 3.8, was less than 50% and decreased further as the pH was increased to 4.3. At the same pH, the recovery of antibodies was greatly increased with 0.5M arginine and more so with 2M arginine. Even at pH 5.0, 2M arginine resulted in 31% recovery, although the elution under such condition showed extensive tailing. Such tailing was observed at pH 3.8 when 0.1M citrate was used. Size exclusion analysis indicated that the eluted antibodies were mostly monomeric whether eluted with citrate or arginine. This demonstrates the usefulness of arginine as an efficient eluent for Protein-A chromatography.

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Citations

May 22, 2013·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hideki WatanabeShinya Honda
Dec 18, 2013·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Sean Wang, Anil Raghani
Jun 28, 2008·Journal of Biochemistry·Atsushi HiranoKentaro Shiraki
Feb 1, 2011·Journal of Biochemistry·Ryosuke ArikiKentaro Shiraki
Feb 9, 2012·Advances in Virology·Hisashi YamasakiA Hajime Koyama
Aug 2, 2011·Biological & Pharmaceutical Bulletin·Tetsuya YoshinoKaori Wakamatsu
Jun 4, 2010·Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan·Tsutomu Arakawa
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Jan 2, 2014·Proceedings of the National Academy of Sciences of the United States of America·Eva-Maria StrauchDavid Baker
Jan 16, 2007·Journal of Pharmaceutical Sciences·Kouhei TsumotoTsutomu Arakawa
Mar 8, 2016·Journal of Chromatography. a·James M Angelo, Abraham M Lenhoff
Jan 12, 2008·Journal of Pharmaceutical Sciences·Hisashi YamasakiTsutomu Arakawa
Nov 7, 2009·Journal of Pharmaceutical Sciences·Tsutomu ArakawaJohn S Philo
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Oct 14, 2008·Protein Expression and Purification·Tsutomu ArakawaDaisuke Ejima
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Aug 30, 2008·Biophysical Chemistry·Makoto NakakidoKouhei Tsumoto
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Apr 6, 2007·Protein Expression and Purification·Tsutomu ArakawaDaisuke Ejima
Jan 30, 2007·Biophysical Chemistry·Tsutomu ArakawaSerge N Timasheff
Nov 28, 2006·Protein Expression and Purification·Tsutomu ArakawaMasao Tokunaga
Aug 19, 2015·Protein Expression and Purification·Tsutomu ArakawaGlen Young
Sep 24, 2009·Biotechnology Progress·Tsuneyoshi MatsuokaKentaro Shiraki
Jan 22, 2009·Biotechnology Journal·Tsutomu ArakawaA Hajime Koyama

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