MEP HyperCel chromatography II: binding, washing and elution

Protein Expression and Purification
Tsutomu ArakawaDaisuke Ejima

Abstract

Binding, washing and elution conditions for MEP HyperCel chromatography were examined using two conditioned media (CM) containing monoclonal antibodies (humanized IgG1) and bovine serum albumin (BSA). Monoclonal antibodies derived from mammalian expression system bound to the column without pretreatment, although a majority of contaminating proteins present in the CM also showed binding. Inorganic salts, ethanol and glycerol were ineffective in eluting proteins under the conditions examined, suggesting that electrostatic or hydrophobic interactions are not a major factor for antibody binding to the MEP resin. Ethylene glycol, 2-propanol, urea and arginine were effective, to varying degrees, in elution of the bound proteins. The bound contaminating proteins and BSA were effectively eluted with ethylene glycol and the bound antibodies were finally eluted with aqueous arginine solutions at neutral pH. MEP showed selectivity toward BSA and hence utility for removing BSA from the samples. Interestingly, Fc-fusion proteins derived from silkworm larvae showed no detectable binding. Serum proteins present in silkworm larvae strongly competed with the Fc-fusion proteins and monoclonal antibody for binding to MEP resin, while the same Fc...Continue Reading

References

Oct 14, 2008·Protein Expression and Purification·Tsutomu ArakawaDaisuke Ejima
Oct 10, 2009·Protein Expression and Purification·Ryosuke YumiokaDaisuke Ejima

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Citations

Aug 24, 2013·PloS One·Yui ShikiyaKentaro Shiraki
Dec 2, 2014·Journal of Chromatography. a·Atsushi HiranoTomoshi Kameda
Mar 4, 2015·Langmuir : the ACS Journal of Surfaces and Colloids·Fang ChengJing-Ping Qu
Jul 14, 2011·Biotechnology Journal·Caryn L HeldtRuben G Carbonell
Apr 5, 2014·Journal of Molecular Recognition : JMR·Simon J MountfordMilton T W Hearn
May 27, 2014·Biochimica Et Biophysica Acta·Tsutomu ArakawaDaisuke Ejima
Mar 20, 2014·Journal of Chromatography. a·Atsushi HiranoTomoshi Kameda
Dec 9, 2014·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·J PezziniX Santarelli
Jul 23, 2016·Protein Expression and Purification·Tsutomu ArakawaYoshiko Kita
Nov 8, 2016·International Journal of Biological Macromolecules·Tsutomu ArakawaTeruo Akuta
Sep 10, 2014·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Karl B McCannJoseph Bertolini
Jul 12, 2017·Bioconjugate Chemistry·Nika Kruljec, Tomaž Bratkovič
Oct 27, 2017·Current Protein & Peptide Science·Xavier Santarelli, Charlotte Cabanne
Oct 27, 2017·Current Protein & Peptide Science·Atsushi HiranoTomoshi Kameda
Nov 19, 2017·Current Protein & Peptide Science·Tsutomu ArakawaKentarou Shiraki
Jan 15, 2020·Journal of Chromatography. a·Julie RobinsonSteven M Cramer
May 25, 2018·Langmuir : the ACS Journal of Surfaces and Colloids·Fang ChengJingping Qu
Dec 6, 2016·Advanced Drug Delivery Reviews·Zehra ElgundiVeysel Kayser

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