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open-access-imgOpen AccessA Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents
Author(s)
Brendan Reed,
Laura B. Chopp,
Courtney S. Malo,
Danielle N. Renner,
Virginia P. Van Keulen,
Megan A. Girtman,
Wendy K. Nevala,
Kevin D. Pavelko,
Diana Gil,
Adam G. Schrum,
Aaron J. Johnson,
Larry R. Pease
Publication year2015
Publication title
plos one
Resource typeJournals
PublisherPublic Library of Science
Antigen-specific T cell responses can be visualized using MHC:peptide multimers. In cases where robust T cell controls are not readily available to assess the integrity of multimer reagents prior to analyzing limited sample, the ability to assess the structural integrity of MHC multimers before their use in critical experiments would be useful. We present a method to probe the structural integrity of MHC multimers using antibodies specific for conformational determinants. Beads coated with anti-mouse Ig are incubated with conformation-specific mouse monoclonal antibody and then with fluorescently tagged MHC multimer. The ability of the bead to capture the labeled multimer can be measured semi-quantitatively by flow cytometry. In this manner, the correct folding of MHC multimers can be visualized and batches of multimer can be compared for quality control. Because there are multiple conformational epitopes formed by various molecular interactions among heavy chain, peptide, and β 2 M, this capture assay can assess the fidelity of each aspect of multimer structure, depending on the availability of antibodies. The described approach could be particularly useful for studies using irreplaceable samples, including patient samples collected in clinical trials.
Subject(s)antibody , antigen , biochemistry , bioinformatics , biology , chemistry , computational biology , electrical engineering , engineering , epitope , flow cytometry , folding (dsp implementation) , immunology , major histocompatibility complex , mhc class i , microbiology and biotechnology , monoclonal antibody , multiplex , peptide , structural engineering , structural integrity
Language(s)English
SCImago Journal Rank0.99
H-Index332
ISSN1932-6203
DOI10.1371/journal.pone.0137984

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