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Performance evaluation of Mimetic Ligand™ B14-triazole-FractoAIMs adsorbents for the capture of human monoclonal immunoglobulin G from cell culture feed

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Abstract

The new affinity-type Mimetic Ligand™ B14 was coupled with a 1,2-diaminoethane spacer (2LP) and a [1,2,3]-triazole spacer (TRZ) to three different support media. In addition to the agarose-based PuraBead and the polymethacrylate-type Fractogel, three new polymeric support media were introduced, the FractoAIMs 1, 2, and 3 (FA1, FA2, and FA3). These new FA supports differ in pore size as well as density of epoxide groups. The immobilization of the B14-ligand onto an azide-group-modified surface was performed with a copper (I)-mediated Click reaction. The IgG capture performance was tested for various ligand–spacer support combinations using cell culture feed containing human immunoglobulin G1 (hIgG1). The most promising adsorbent, B14-TRZ-FA3, was further optimized by improving the surface chemistry through a triple endcap** concept employing an improved Click reaction protocol. This new technique enabled the most efficient deactivation of residual azide groups. In a direct comparison with a commercially available Protein A media, B14-TRZ-FA3 3× ec provided superior results at fast flow-rates and low bed-height. Dynamic binding capacities of 11.4 g/L for 10% breakthrough of hIgG1, elution capacities of 16.0 g/L hIgG1 and a recovery of 86% were achieved. The same results were obtained for a dialyzed and pre-purified feed solution, which is a clear indicator that triple-endcapped affinity support surfaces are practically inert to the non-specific binding of host cell proteins.

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Acknowledgments

This study was performed within the EU-Project AIMs (Advanced Interactive Materials by Design; NMP3-CT-2004-500160), which is part of the 6th Framework Program of the European Union. The authors want to thank Merck KGaA for providing the Fractogel® and FractoAIMs® support media. Note that ProMetic BioSciences and Mimetic Ligands™ are trademarks of ProMetic BioSciences Ltd and that PuraBead® is registered with the U.S. Patent & Trademark Office.

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Authors and Affiliations

  1. Department of Analytical Chemistry, University of Vienna, Waehringerstrasse 38, 1090, Vienna, Austria

    Jeannie Horak, Stefan Hofer & Wolfgang Lindner

  2. ProMetic BioSciences Ltd., 211 Cambridge Science Park, Milton Road, Cambridge, CB4 0WA, UK

    Chris Sadler & Sharon Williams

Authors
  1. Jeannie Horak
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  2. Stefan Hofer
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  3. Chris Sadler
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  4. Sharon Williams
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  5. Wolfgang Lindner
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Correspondence to Jeannie Horak.

Additional information

Published in the special issue Analytical Sciences in Austria with Guest Editors G. Allmaier, W. Buchberger, and K. Francesconi.

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Horak, J., Hofer, S., Sadler, C. et al. Performance evaluation of Mimetic Ligand™ B14-triazole-FractoAIMs adsorbents for the capture of human monoclonal immunoglobulin G from cell culture feed. Anal Bioanal Chem 400, 2349–2359 (2011). https://doi.org/10.1007/s00216-010-4571-1

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  • DOI: https://doi.org/10.1007/s00216-010-4571-1

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