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Entrapment of Proteins Within Columns for High-Performance Affinity Chromatography

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Affinity Chromatography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2466))

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Abstract

Entrapment is a noncovalent immobilization method that enables a large biological binding agent, such as a protein, to be put within a support without modifying the structure of the binding agent. This chapter describes an on-column entrapment method that can be used with proteins and HPLC-grade silica to prepare columns for high-performance liquid chromatography. In this method, a protein is trapped within a dihydrazide-activated silica support by using oxidized glycogen as a capping agent. This method allows the protein to be placed within the support in a soluble form and with little or no loss of activity. The approach and reagents needed for this method are described in this chapter, along with some applications reported for columns that have been made using on-column protein entrapment.

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Acknowledgments

This work was supported, in part, by the University of Nebraska Research Council, the University of Nebraska-Lincoln College of Arts and Sciences, the National Science Foundation under grant CHE 2108881, and the National Institutes of Health under grant R01 DK069629.

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Authors and Affiliations

  1. Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE, USA

    Saumen Poddar, Sadia Sharmeen & David S. Hage

Authors
  1. Saumen Poddar
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  2. Sadia Sharmeen
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  3. David S. Hage
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Corresponding author

Correspondence to David S. Hage .

Editor information

Editors and Affiliations

  1. Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, USA

    B. Vijayalakshmi Ayyar

  2. Astero Erado Inc, Houston, TX, USA

    Sushrut Arora

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Poddar, S., Sharmeen, S., Hage, D.S. (2022). Entrapment of Proteins Within Columns for High-Performance Affinity Chromatography. In: Ayyar, B.V., Arora, S. (eds) Affinity Chromatography. Methods in Molecular Biology, vol 2466. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2176-9_14

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  • DOI: https://doi.org/10.1007/978-1-0716-2176-9_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2175-2

  • Online ISBN: 978-1-0716-2176-9

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