Nature Biotechnology22, 1429 - 1434 (2004)
Published online: 24 October 2004; | doi:10.1038/nbt1024
Engineered T-cell receptor tetramers bind MHC-peptide complexes with high affinity
Ramu A Subbramanian1, 3, Chikaya Moriya1, 3, Kristi L Martin1, 3, Fred W Peyerl1, Atsuhiko Hasegawa1, Akira Naoi1, Heng Chhay1, Patrick Autissier1, Darci A Gorgone1, Michelle A Lifton1, Kristine Kuus-Reichel2, Jörn E Schmitz1, Norman L Letvin1
& Marcelo J Kuroda1
1
Division of Viral Pathogenesis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, Massachusetts 02215, USA.
2
Immunomics Operations, Beckman Coulter, San Diego, California 92121, USA.
In this study we extend tetramerization technology to T-cell receptors (TCRs). We identified TCR pairs in the absence of accessory molecules, ensuring isolation of high-affinity TCRs that maintain stable binding characteristics after tetramerization. Subtle changes in cognate peptide levels bound to the class I molecule were accurately reflected by parallel changes in the mean fluorescence intensity of cells that bound TCR tetramers, allowing us to accurately assess the binding affinity of a panel of peptides to major histocompatibility complex (MHC) class I. Using a TCR tetramer specific for the Mamu-A*01 allele, we identified animals expressing this restricting class I allele from a large cohort of outbred rhesus macaques. TCR tetramers should facilitate analysis of the MHC-peptide interface and, more generally, the design of immunotherapeutics and vaccines.
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pairs in the absence of accessory molecules, ensuring isolation of high-affinity TCRs that maintain stable binding characteristics after tetramerization. Subtle changes in cognate peptide levels bound to the class I molecule were accurately reflected by parallel changes in the mean fluorescence intensity of cells that bound TCR tetramers, allowing us to accurately assess the binding affinity of a panel of peptides to major histocompatibility complex (MHC) class I. Using a TCR tetramer specific for the Mamu-A*01 allele, we identified animals expressing this restricting class I allele from a large cohort of outbred rhesus macaques. TCR tetramers should facilitate analysis of the MHC-peptide interface and, more generally, the design of immunotherapeutics and vaccines.
