Summary
High-throughput protein purification is a complex, multi-step process. There are several technical challenges in the course of this process that are not experienced when purifying a single protein. Among the most challenging are the high-throughput protein concentration and buffer exchange, which are not only labor-intensive but can also result in significant losses of purified proteins. We describe two methods of high-throughput protein concentration and buffer exchange: one using ammonium sulfate precipitation and one using micro-concentrating devices based on membrane ultrafiltration. We evaluated the efficiency of both methods on a set of 18 randomly selected purified proteins from Shewanella oneidensis. While both methods provide similar yield and efficiency, the ammonium sulfate precipitation is much less labor intensive and time consuming than the ultrafiltration.
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Acknowledgments
This work was supported by the U.S. Department of Energy Genomics: GTL Program, Grant #KP1 102010. This manuscript has been authored by Battelle Memorial Institute, Pacific Northwest Division, under Contract No. DE-AC05—76RL0 1830 with the U.S. Department of Energy. The publisher, by accepting the article for publication, acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Moore, P.A., Kery, V. (2009). High-Throughput Protein Concentration and Buffer Exchange: Comparison of Ultrafiltration and Ammonium Sulfate Precipitation. In: Doyle, S.A. (eds) High Throughput Protein Expression and Purification. Methods in Molecular Biology, vol 498. Humana Press. https://doi.org/10.1007/978-1-59745-196-3_20
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DOI: https://doi.org/10.1007/978-1-59745-196-3_20
Publisher Name: Humana Press
Print ISBN: 978-1-58829-879-9
Online ISBN: 978-1-59745-196-3
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