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Major histocompatibility complex (MHC) class I multimer technology has become an indispensable immunological assay system to dissect antigen-specific cytotoxic CD8(+) T cell responses by flow cytometry. However, the development of high-throughput assay systems, in which T cell responses against a multitude of epitopes are analyzed, has been precluded by the fact that for each T cell epitope, a separate in vitro MHC refolding reaction is required. We have recently demonstrated that conditional ligands that disintegrate upon exposure to long-wavelength UV light can be designed for the human MHC molecule HLA-A2. To determine whether this peptide-exchange technology can be developed into a generally applicable approach for high throughput MHC based applications we set out to design conditional ligands for the human MHC gene products HLA-A1, -A3, -A11, and -B7. Here, we describe the development and characterization of conditional ligands for this set of human MHC molecules and apply the peptide-exchange technology to identify melanoma-associated peptides that bind to HLA-A3 with high affinity. The conditional ligand technology developed here will allow high-throughput MHC-based analysis of cytotoxic T cell immunity in the vast majority of Western European individuals.

Original publication




Journal article


Proceedings of the National Academy of Sciences of the United States of America

Publication Date





3825 - 3830


Divisions of Immunology and Cellular Biochemistry, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.


CD8-Positive T-Lymphocytes, Clone Cells, Humans, Melanoma, Peptides, Histocompatibility Antigens Class I, HLA-A Antigens, HLA-A1 Antigen, HLA-A3 Antigen, HLA-B7 Antigen, Epitopes, Ligands, Protein Engineering, Inhibitory Concentration 50, Ultraviolet Rays, Protein Structure, Quaternary, Protein Folding, Kinetics, Alleles, HLA-A11 Antigen