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Two Elution Mechanisms of MEP Chromatography

[ Vol. 20 , Issue. 1 ]

Author(s):

Tsutomu Arakawa*, Masao Tokunaga, Takuya Maruyama and Kentaro Shiraki   Pages 28 - 33 ( 6 )

Abstract:


MEP (mercapto-ethyl-pyridine) HyperCel is one of the hydrophobic charge induction chromatography (HCIC) resins. Under normal operation, proteins are bound to the MEP resin at neutral pH, at which MEP is not charged, mostly via hydrophobic interaction. MEP has a pyridine group, whose pK is 4.8, and hence is positively charged at acidic pH range. Based on the binding mechanism (i.e., hydrophobic interaction) and the induced positive charge at acidic pH, there may be two ways to elute the bound proteins. One way is to bring the pH down to protonate both MEP resin and the bound protein, leading to charge repulsion and thereby elution. Another way is to use hydrophobic interaction modifiers, which are often used in hydrophobic interaction chromatography, to reduce hydrophobic interaction. Here, we summarize such two possible elution approaches.

Keywords:

MEP HyperCel, chromatography, antibody, arginine, elution, acidic pH.

Affiliation:

Alliance Protein Laboratories, A Division of KBI Biopharma, 6042 Cornerstone Court West, San Diego, CA 92121, Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Faculty of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305- 8573, Faculty of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305- 8573

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