Competition-based cellular peptide binding assays for 13 prevalent HLA class I alleles using fluorescein-labeled synthetic peptides

Jan H. Kessler*, Bregje Mommaas, Tuna Mutis, Ivo Huijbers, Debby Vissers, Willemien E. Benckhuijsen, Geziena M.Th Schreuder, Rienk Offringa, Els Goulmy, Cornelis J.M. Melief, Sjoerd H. Van der Burg, Jan W. Drijfhout

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

We report the development, validation, and application of competition-based peptide binding assays for 13 prevalent human leukocyte antigen (HLA) class I alleles. The assays are based on peptide binding to HLA molecules on living cells carrying the particular allele. Competition for binding between the test peptide of interest and a fluorescein-labeled HLA class I binding peptide is used as read out. The use of cell membranebound HLA class I molecules circumvents the need for laborious biochemical purification of these molecules in soluble form. Previously, we have applied this principle for HLA-A2 and HLA-A3. We now describe the assays for HLA-A1, HLA-A11, HLA-A24, HLA-A68, HLA-B7, HLA-B8, HLA-B14, HLA-B35, HLA-B60, HLA-B61, and HLA-B62. Together with HLA-A2 and HLA-A3, these alleles cover more than 95% of the Caucasian pop ulation. Several allele-specific parameters were determined for each assay. Using these assays, we identified novel HLA class I high-affinity binding peptides from HIVpol, p53, PRAME, and minor histocompatibility antigen HA-1. Thus these convenient and accurate peptidebinding assays will be useful for the identification of putative cytotoxic T lymphocyte epitopes presented on a diverse array of HLA class I molecules.

Original languageEnglish
Pages (from-to)245-255
Number of pages11
JournalHuman Immunology
Volume64
Issue number2
DOIs
Publication statusPublished - 1 Feb 2003

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