High-throughput T-cell epitope discovery through MHC peptide exchange

SR Hadrup, M Toebes, B Rodenko, AH Bakker… - Epitope Mapping …, 2009 - Springer
SR Hadrup, M Toebes, B Rodenko, AH Bakker, DA Egan, H Ovaa, TNM Schumacher
Epitope Mapping Protocols: Second Edition, 2009Springer
Recombinant major histocompatibility complex (MHC) class I molecules complexed with
pathogen-specific or other disease-associated antigens have become essential reagents for
the analysis of adaptive T-cell responses. However, conventional techniques for the
production of recombinant peptide-MHC (pMHC) complexes are highly involved and thereby
limit the use of pMHC complexes in terms of antigen diversity. To make pMHC-based
techniques suitable for high-throughput analyses we developed an MHC peptide exchange …
Summary
Recombinant major histocompatibility complex (MHC) class I molecules complexed with pathogen-specific or other disease-associated antigens have become essential reagents for the analysis of adaptive T-cell responses. However, conventional techniques for the production of recombinant peptide-MHC (pMHC) complexes are highly involved and thereby limit the use of pMHC complexes in terms of antigen diversity. To make pMHC-based techniques suitable for high-throughput analyses we developed an MHC peptide exchange technology based on the use of conditional MHC ligands. This technology enables the parallel production of thousands of different pMHC complexes within hours, allowing the development of high-throughput MHC-based assay systems to identify MHC ligands and cytotoxic T-cell responses. These high-throughput assays should prove valuable for the screening of entire disease-associated proteomes, including pathogen-encoded proteomes, tumor-associated antigens, and autoimmune antigens.
Springer