title: A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents.
abstract: 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 β2M, 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.
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Brendan K
authors.initials: BK
authors.lastname: Reed
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Laura B
authors.initials: LB
authors.lastname: Chopp
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Courtney S
authors.initials: CS
authors.lastname: Malo
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Danielle N
authors.initials: DN
authors.lastname: Renner
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Virginia S
authors.initials: VS
authors.lastname: Van Keulen
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Megan A
authors.initials: MA
authors.lastname: Girtman
authors.affiliation: Department of Medicine, Division of Hematology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Wendy N
authors.initials: WN
authors.lastname: Nevala
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Pavelko
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Diana
authors.initials: D
authors.lastname: Gil
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Adam G
authors.initials: AG
authors.lastname: Schrum
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Johnson
authors.affiliation: Department of Immunology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester, MN, United States of America.
authors.forename: Larry R
authors.initials: LR
authors.lastname: Pease
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology*; D051379:Mice; D011487:Protein Conformation; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000941:Antigens