A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents.
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 thei...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4577079?pdf=render |
| _version_ | 1818848185289801728 |
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| author | Brendan K Reed Laura B Chopp Courtney S Malo Danielle N Renner Virginia S Van Keulen Megan A Girtman Wendy N Nevala Kevin D Pavelko Diana Gil Adam G Schrum Aaron J Johnson Larry R Pease |
| author_facet | Brendan K Reed Laura B Chopp Courtney S Malo Danielle N Renner Virginia S Van Keulen Megan A Girtman Wendy N Nevala Kevin D Pavelko Diana Gil Adam G Schrum Aaron J Johnson Larry R Pease |
| author_sort | Brendan K Reed |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-19T06:13:19Z |
| format | Article |
| id | doaj.art-1fb448f2255f452cad3fed8f44b99cae |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-19T06:13:19Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-1fb448f2255f452cad3fed8f44b99cae2022-12-21T20:32:56ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01109e013798410.1371/journal.pone.0137984A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents.Brendan K ReedLaura B ChoppCourtney S MaloDanielle N RennerVirginia S Van KeulenMegan A GirtmanWendy N NevalaKevin D PavelkoDiana GilAdam G SchrumAaron J JohnsonLarry R PeaseAntigen-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.http://europepmc.org/articles/PMC4577079?pdf=render |
| spellingShingle | Brendan K Reed Laura B Chopp Courtney S Malo Danielle N Renner Virginia S Van Keulen Megan A Girtman Wendy N Nevala Kevin D Pavelko Diana Gil Adam G Schrum Aaron J Johnson Larry R Pease A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents. PLoS ONE |
| title | A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents. |
| title_full | A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents. |
| title_fullStr | A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents. |
| title_full_unstemmed | A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents. |
| title_short | A Versatile Simple Capture Assay for Assessing the Structural Integrity of MHC Multimer Reagents. |
| title_sort | versatile simple capture assay for assessing the structural integrity of mhc multimer reagents |
| url | http://europepmc.org/articles/PMC4577079?pdf=render |
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