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An exclusion mechanism in ion exchange chromatography
Author(s) -
Harinarayan C.,
Mueller J.,
Ljunglöf A.,
Fahrner R.,
Van Alstine J.,
van Reis R.
Publication year - 2006
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21080
Subject(s) - conductivity , chemistry , ion chromatography , ion , ion exchange , mechanism (biology) , biophysics , chromatography , chemical engineering , chemical physics , analytical chemistry (journal) , organic chemistry , biology , engineering , philosophy , epistemology
Protein dynamic binding capacities on ion exchange resins are typically expected to decrease with increasing conductivity and decreasing protein charge. There are, however, conditions where capacity increases with increasing conductivity and decreasing protein charge. Capacity measurements on two different commercial ion exchange resins with three different monoclonal antibodies at various pH and conductivities exhibited two domains. In the first domain, the capacity unexpectedly increased with increasing conductivity and decreasing protein charge. The second domain exhibited traditional behavior. A mechanism to explain the first domain is postulated; proteins initially bind to the outer pore regions and electrostatically hinder subsequent protein transport. Such a mechanism is supported by protein capacity and confocal microscopy studies whose results suggest how knowledge of the two types of IEX behavior can be leveraged in optimizing resins and processes. © 2006 Wiley Periodicals, Inc.