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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 681))

Abstract

Affinity chromatography permits the isolation of a target analyte from a complex mixture and can be utilised 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 proteins 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 proteins 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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Acknowledgements

This material is based upon works supported by the Science Foundation Ireland under Grant No. 05/CE3/B754 and the Food Institutional Research Measure (FIRM) Department of Agriculture, under Grant No. 05/RND/TN355.

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Correspondence to Richard O’Kennedy .

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Darcy, E., Leonard, P., Fitzgerald, J., Danaher, M., O’Kennedy, R. (2011). Purification of Antibodies Using Affinity Chromatography. 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_20

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  • DOI: https://doi.org/10.1007/978-1-60761-913-0_20

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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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