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dc.contributor.authorYin, Liusong
dc.contributor.authorStern, Lawrence J.
dc.date2022-08-11T08:08:55.000
dc.date.accessioned2022-08-23T16:12:20Z
dc.date.available2022-08-23T16:12:20Z
dc.date.issued2014-04-01
dc.date.submitted2015-08-31
dc.identifier.citationJ Immunol Methods. 2014 Apr;406:21-33. doi: 10.1016/j.jim.2014.02.008. Epub 2014 Feb 25. <a href="http://dx.doi.org/10.1016/j.jim.2014.02.008">Link to article on publisher's site</a>
dc.identifier.issn0022-1759 (Linking)
dc.identifier.doi10.1016/j.jim.2014.02.008
dc.identifier.pmid24583195
dc.identifier.urihttp://hdl.handle.net/20.500.14038/33374
dc.description.abstractHLA-DM (DM) functions as a peptide editor that mediates the exchange of peptides loaded onto MHCII molecules by accelerating peptide dissociation and association kinetics. The relative DM-susceptibility of peptides bound to MHCII molecules correlates with antigen presentation and immunodominance hierarchy, and measurement of DM-susceptibility has been a key effort in this field. Current assays of DM-susceptibility, based on differential peptide dissociation rates measured for individually labeled peptides over a long time base, are difficult and cumbersome. Here, we present a novel method to measure DM-susceptibility based on peptide binding competition assays performed in the presence and absence of DM, reported as a delta-IC(50) (change in 50% inhibition concentration) value. We simulated binding competition reactions of peptides with various intrinsic and DM-catalyzed kinetic parameters and found that under a wide range of conditions the delta-IC(50) value is highly correlated with DM-susceptibility as measured in off-rate assay. We confirmed experimentally that DM-susceptibility measured by delta-IC(50) is comparable to that measured by traditional off-rate assay for peptides with known DM-susceptibility hierarchy. The major advantage of this method is that it allows simple, fast and high throughput measurement of DM-susceptibility for a large set of unlabeled peptides in studies of the mechanism of DM action and for identification of CD4+ T cell epitopes.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=24583195&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC4029850/
dc.subjectAntibody Affinity; Antigen Presentation; Binding, Competitive; CD4-Positive T-Lymphocytes; Epitopes, T-Lymphocyte; Fluorescence Polarization; HLA-D Antigens; HLA-DR alpha-Chains; HLA-DRB1 Chains; Hemagglutinin Glycoproteins, Influenza Virus; Histocompatibility Antigens Class II; Humans; Inhibitory Concentration 50; Peptide Fragments; Protein Binding
dc.subjectBiochemistry
dc.subjectImmunology and Infectious Disease
dc.subjectImmunopathology
dc.titleA novel method to measure HLA-DM-susceptibility of peptides bound to MHC class II molecules based on peptide binding competition assay and differential IC(50) determination
dc.typeJournal Article
dc.source.journaltitleJournal of immunological methods
dc.source.volume406
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/gsbs_sp/1899
dc.identifier.contextkey7536272
html.description.abstract<p>HLA-DM (DM) functions as a peptide editor that mediates the exchange of peptides loaded onto MHCII molecules by accelerating peptide dissociation and association kinetics. The relative DM-susceptibility of peptides bound to MHCII molecules correlates with antigen presentation and immunodominance hierarchy, and measurement of DM-susceptibility has been a key effort in this field. Current assays of DM-susceptibility, based on differential peptide dissociation rates measured for individually labeled peptides over a long time base, are difficult and cumbersome. Here, we present a novel method to measure DM-susceptibility based on peptide binding competition assays performed in the presence and absence of DM, reported as a delta-IC(50) (change in 50% inhibition concentration) value. We simulated binding competition reactions of peptides with various intrinsic and DM-catalyzed kinetic parameters and found that under a wide range of conditions the delta-IC(50) value is highly correlated with DM-susceptibility as measured in off-rate assay. We confirmed experimentally that DM-susceptibility measured by delta-IC(50) is comparable to that measured by traditional off-rate assay for peptides with known DM-susceptibility hierarchy. The major advantage of this method is that it allows simple, fast and high throughput measurement of DM-susceptibility for a large set of unlabeled peptides in studies of the mechanism of DM action and for identification of CD4+ T cell epitopes.</p>
dc.identifier.submissionpathgsbs_sp/1899
dc.contributor.departmentDepartment of Biochemistry and Molecular Pharmacology
dc.contributor.departmentDepartment of Pathology
dc.contributor.departmentProgram in Immunology and Microbiology
dc.source.pages21-33
dc.contributor.studentLiusong Yin


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