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Measuring Antibody-Antigen Binding Kinetics Using Surface Plasmon Resonance

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Antibody Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1827))

Abstract

Surface plasmon resonance (SPR) is now widely embraced as a technology for monitoring a diverse range of protein-protein interactions and is considered almost de rigueur for characterizing antibody-antigen interactions. The technique obviates the need to label either of the interacting species, and the binding event is visualized in real time. Thus, it is ideally suited for screening crude, unpurified antibody samples that dominate early candidate panels following antibody selection campaigns. SPR returns not only concentration and affinity data but when used correctly can resolve the discrete component kinetic parameters (association and dissociation rate constants) of the affinity interaction. Herein, we outline some SPR-based generic antibody screening configurations and methodologies in the context of expediting data-rich ranking of candidate antibody panels and ensuring that antibodies with the optimal kinetic binding characteristics are reliably identified.

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Acknowledgments

The authors gratefully acknowledge the technical contributions from C. J. Hayes, P. J. Conroy, and B. Vijayalakshmi Ayyar. This material is based on work supported by the Science Foundation Ireland (grants 10/CE/B1821 and 14/IA/2646) and Enterprise Ireland (grant CF/2015/0105).

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Authors and Affiliations

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

    Stephen Hearty, Paul Leonard, Hui Ma & Richard O’Kennedy

  2. National Centre for Sensor Research, Dublin City University, Dublin, Ireland

    Stephen Hearty, Paul Leonard & Richard O’Kennedy

  3. Qatar Foundation and Research Complex, Hamad Bin Khalifa University, Education City, Doha, Qatar

    Richard O’Kennedy

Authors
  1. Stephen Hearty
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  2. Paul Leonard
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  3. Hui Ma
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  4. Richard O’Kennedy
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Corresponding author

Correspondence to Richard O’Kennedy .

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Editors and Affiliations

  1. Immunology Division, Garvan Institute of Medical Research, Darlinghurst, NSW, Australia

    Damien Nevoltris

  2. Institut Paoli-Calmettes, CRCM, Aix Marseille University, CNRS, INSERM, Marseille, France

    Patrick Chames

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Hearty, S., Leonard, P., Ma, H., O’Kennedy, R. (2018). Measuring Antibody-Antigen Binding Kinetics Using Surface Plasmon Resonance. In: Nevoltris, D., Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, vol 1827. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8648-4_22

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  • DOI: https://doi.org/10.1007/978-1-4939-8648-4_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8647-7

  • Online ISBN: 978-1-4939-8648-4

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