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Purification of Antibodies Using Affinity Chromatography

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Protein Chromatography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1485))

Abstract

Affinity chromatography permits the isolation of a target analyte from a complex mixture and can be utilized to purify proteins, carbohydrates, drugs, haptens, or any analyte of interest once an affinity pair is available. It involves the exploitation of specific interactions between a binding affinity pair, such as those between an antibody and its associated antigen, or between any ligand and its associated binding receptor/protein. With the discovery of protein A in 1970, and, subsequently protein G and L, immuno-affinity chromatography has grown in popularity and is now the standard methodology for the purification of antibodies which may be implemented for a selection of different applications such as immunodiagnostics. This chapter is designed to inform the researcher about the basic techniques involved in the affinity chromatography-based purification of monoclonal, polyclonal, and recombinant antibodies. Examples are provided for the use of protein A and G. In addition, tables are provided that allow the reader to select the most appropriate protein for use in the isolation of their antibody.

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Acknowledgments

This material is based on works supported by Science Foundation Ireland (Grants 05/CE3/B754 and 14/IA/2646), the Food Institutional Research Measure (FIRM) of the Department of Agriculture, Food and the Marine of Ireland (Grant 05/RND/TN355) and Enterprise Ireland (Grant CF/2015/0105P).

Author information

Authors and Affiliations

  1. School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland

    Elaine Darcy, Paul Leonard, Jenny Fitzgerald & Richard O’Kennedy

  2. Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland

    Paul Leonard, Hui Ma & Richard O’Kennedy

  3. Ashtown Food Research Centre, Teagasc, Ashtown, Dublin 15, Ireland

    Martin Danaher

Authors
  1. Elaine Darcy
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  2. Paul Leonard
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  3. Jenny Fitzgerald
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  4. Martin Danaher
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  5. Hui Ma
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  6. Richard O’Kennedy
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Corresponding author

Correspondence to Richard O’Kennedy .

Editor information

Editors and Affiliations

  1. School of Biotechnology, Dublin City University, Dublin, Ireland

    Dermot Walls

  2. Department of Applied Sciences, Dundalk Institute of Technology, Dublin, Ireland

    Sinéad T. Loughran

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Darcy, E., Leonard, P., Fitzgerald, J., Danaher, M., Ma, H., O’Kennedy, R. (2017). Purification of Antibodies Using Affinity Chromatography. In: Walls, D., Loughran, S. (eds) Protein Chromatography. Methods in Molecular Biology, vol 1485. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6412-3_15

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  • DOI: https://doi.org/10.1007/978-1-4939-6412-3_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6410-9

  • Online ISBN: 978-1-4939-6412-3

  • eBook Packages: Springer Protocols

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