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Biotechnology and Applied Biochemistry (2010) 56, (59–70) (Printed in Great Britain)

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Overloading ion-exchange membranes as a purification step for monoclonal antibodies

Arick Brown¹, Jerome Bill, Timothy Tully, Asha Radhamohan and Chris Dowd

Process Technical Development, Genentech, 1 DNA Way, South San Francisco, CA 94080, U.S.A.

Key words: competitive adsorption, displacement chromatography, flow-through chromatography, ion-exchange membrane, monoclonal antibody (mAb), overload chromatography.

Abbreviations used: CEX, cation exchange; CHO, Chinese-hamster ovary; CHOP, CHO protein; HRP, horseradish peroxidase; LOQ, limit of quantification; mAb, monoclonal antibody; MV, membrane volume; PPM, parts per million; SEC, size-exclusion chromatography.

¹To whom correspondence should be addressed (email abrown@gene.com).

The present study examined the overloading of ion-exchange membrane adsorbers, a form of frontal chromatography, as the final purification step in the production of mAbs (monoclonal antibodies) produced from CHO (Chinese-hamster ovary) cells. Preferential binding of impurities over antibody product was exploited using commercially available cation- and anion-exchange membranes. Three different antibody feedstreams previously purified over Protein A and ion-exchange column chromatography were tested. Feedstream conductivity and pH were adjusted to induce product and impurity adsorption. Membranes were then overloaded in a normal flow mode, resulting in retention of impurities and breakthrough of purified antibody. Although some amount of the product also binds to the membranes (usually ≤30 g mAb/l membrane), yields of ≥99% were achieved by marginalizing the losses, typically by loading more than 3 kg mAb/l membrane. Analyses of the purified pools show consistent removal of impurities despite strong mAb–ligand interactions and high membrane loadings. The clearance of host cell proteins was affected by pH and conductivity, but was unaffected by flow rate, membrane properties or scale. The importance of the present study lies in our demonstration of an alternative use of ion-exchange membranes for fast, effective and high yielding purification of mAbs.

Received 23 December 2009/6 May 2010; accepted 19 May 2010

Published as Immediate Publication 19 May 2010, doi:10.1042/BA20090369

© 2010 The Author(s)
The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Non-Commercial Licence (http://creativecommons.org/licenses/by-nc/2.5/) which permits unrestricted non-commercial use, distribution and reproduction in any medium, provided the original work is properly cited.