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Ultra-Performance Liquid Chromatography–Mass Spectrometry of Proteins

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Protein Chromatography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 681))

Abstract

The commercialization of ultra-performance liquid chromatography (UPLC) has allowed more researchers to take advantage of the benefits of this work. Many researchers are exploring this technique to reduce analytical throughput and to increase resolution. The majority of this work has focused on small molecule analysis; however, this technique can provide the same advantages for the analysis of proteins. Traditionally, protein chromatography has suffered from a number of issues such as carryover, peak splitting, peak broadening, and poor peak shape. Because UPLC utilizes a smaller particle at a higher pressure and flow rate, many of these issues are remedied. When used in combination with mass spectrometry, UPLC becomes a powerful tool for protein identification, characterization, and quantitation. In this work we show how UPLC/MS can be used to separate and identify intact proteins.

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Acknowledgments

The author would like to thank Robert A. Everley, Ph.D. for his contributions to this work. A portion of this work was funded by the Centers for Disease Control and Prevention (CDC) grant # U90/CCU317014.

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Correspondence to Timothy R. Croley .

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Croley, T.R. (2011). Ultra-Performance Liquid Chromatography–Mass Spectrometry of Proteins. In: Walls, D., Loughran, S. (eds) Protein Chromatography. Methods in Molecular Biology, vol 681. Humana Press. https://doi.org/10.1007/978-1-60761-913-0_23

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  • DOI: https://doi.org/10.1007/978-1-60761-913-0_23

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-912-3

  • Online ISBN: 978-1-60761-913-0

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