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Isolation of Major Histocompatibility Complex (MHC)-Associated Peptides by Immunoaffinity Purification

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Immunoproteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2024))

Abstract

MHC molecules on the surface of cells are responsible for the presentation of antigenic peptides to CD8+ and CD4+ T cells. Downstream analysis of these peptides can offer insight into various disease processes with an immune component, as in autoimmune diseases and cancer. A critical step in studying MHC-associated peptides is their isolation from MHC molecules on the surface of cells. In this chapter, we detail anti-MHC antibody conjugation to beads, immunoaffinity purification of the MHC molecules, and peptide elution. This method can be used for analysis of unmodified and/or posttranslationally modified peptides by mass spectrometry.

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Acknowledgments

The authors would like to acknowledge Mark Cobbold (MGH) and Donald F. Hunt (U Virginia) for their technical and financial support. S.A.M. is currently funded by an NIH F32 postdoctoral fellowship.

Author information

Authors and Affiliations

  1. Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, UK

    Sarah A. Penny

  2. Department of Chemistry, Stanford University, Stanford, CA, USA

    Stacy A. Malaker

Authors
  1. Sarah A. Penny
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  2. Stacy A. Malaker
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Corresponding author

Correspondence to Stacy A. Malaker .

Editor information

Editors and Affiliations

  1. Human Health Therapeutics Research Centre, National Research Council of Canada, Ottawa, ON, Canada

    Kelly M. Fulton

  2. Human Health Therapeutics Research Centre, National Research Council of Canada, Ottawa, ON, Canada

    Susan M. Twine

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© 2019 Springer Science+Business Media, LLC, part of Springer Nature

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Penny, S.A., Malaker, S.A. (2019). Isolation of Major Histocompatibility Complex (MHC)-Associated Peptides by Immunoaffinity Purification. In: Fulton, K., Twine, S. (eds) Immunoproteomics. Methods in Molecular Biology, vol 2024. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9597-4_14

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  • DOI: https://doi.org/10.1007/978-1-4939-9597-4_14

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9596-7

  • Online ISBN: 978-1-4939-9597-4

  • eBook Packages: Springer Protocols

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