Description
This protocol is designed to evaluate the efficiency of peptide-HLA-I interaction and complex formation. The assay is based on detecting the ß2-microglobulin (ß2m? light chain subunit of recombinant HLA Class I (HLA-I) comlexes, where the heavy chain has been biotin tagged. These tagged complexes are subsequently captured by streptavidin coated beads, labeled with PE-conjugated anti-human ß2m, and analyzed by flow cytometry. Since peptide-HLA-I complex formation is entirely peptide dependent, bead-associated signals will only be detected if the peptide in question supports the folding of the HLA-I allotype of interest; peptides that efficiently support folding will give strong signals whereas peptides that support folding sub-optimally, or not at all, will give moderate to non-detectable signals