GSBS Student Publications
Title
Measurement of Peptide Binding to MHC Class II Molecules by Fluorescence Polarization
GSBS Program
Immunology & Microbiology
Publication Date
2014-08-01
UMMS Affiliation
Department of Pathology; Department of Biochemistry and Molecular Pharmacology
Document Type
Article
Disciplines
Biochemistry | Immunology and Infectious Disease | Immunology of Infectious Disease
Abstract
Peptide binding to major histocompatibility complex class II (MHCII) molecules is a key process in antigen presentation and CD4+ T cell epitope selection. This unit describes a fairly simple but powerful fluorescence polarization-based binding competition assay to measure peptide binding to soluble recombinant MHCII molecules. The binding of a peptide of interest to MHCII molecules is assessed based on its ability to inhibit the binding of a fluorescence-labeled probe peptide, with the strength of binding characterized as IC50 (concentration required for 50% inhibition of probe peptide binding). Data analysis related to this method is discussed. In addition, this unit includes a support protocol for fluorescence labeling peptide using an amine-reactive probe. The advantage of this protocol is that it allows simple, fast, and high-throughput measurements of binding for a large set of peptides to MHCII molecules.
DOI of Published Version
10.1002/0471142735.im0510s106
Source
Curr Protoc Immunol. 2014 Aug 1;106:5.10.1-5.10.12. doi: 10.1002/0471142735.im0510s106. Link to article on publisher's site
Journal/Book/Conference Title
Current protocols in immunology / edited by John E. Coligan ... [et al.]
Related Resources
PubMed ID
25081912
Repository Citation
Yin L, Stern LJ. (2014). Measurement of Peptide Binding to MHC Class II Molecules by Fluorescence Polarization. GSBS Student Publications. https://doi.org/10.1002/0471142735.im0510s106. Retrieved from https://escholarship.umassmed.edu/gsbs_sp/1930