title: Directed evolution of human T-cell receptors with picomolar affinities by phage display.
abstract: Peptides derived from almost all proteins, including disease-associated proteins, can be presented on the cell surface as peptide-human leukocyte antigen (pHLA) complexes. T cells specifically recognize pHLA with their clonally rearranged T-cell receptors (TCRs), whose natural affinities are limited to approximately 1-100 muM. Here we describe the display of ten different human TCRs on the surface of bacteriophage, stabilized by a nonnative interchain disulfide bond. We report the directed evolution of high-affinity TCRs specific for two different pHLAs: the human T-cell lymphotropic virus type 1 (HTLV-1) tax(11-19) peptide-HLA-A(*)0201 complex and the NY-ESO-1(157-165) tumor-associated peptide antigen-HLA-A(*)0201 complex, with affinities of up to 2.5 nM and 26 pM, respectively, and we demonstrate their high specificity and sensitivity for targeting of cell-surface pHLAs.
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.forename: Ruth
authors.initials: R
authors.lastname: Moysey
authors.forename: Peter E
authors.initials: PE
authors.lastname: Molloy
authors.forename: Anne-Lise
authors.initials: AL
authors.lastname: Vuidepot
authors.forename: Tara
authors.initials: T
authors.lastname: Mahon
authors.forename: Emma
authors.initials: E
authors.lastname: Baston
authors.forename: Steven
authors.initials: S
authors.lastname: Dunn
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Liddy
authors.forename: Jansen
authors.initials: J
authors.lastname: Jacob
authors.forename: Bent K
authors.initials: BK
authors.lastname: Jakobsen
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Boulter
mesh_terms: D000915:Antibody Affinity; D000917:Antibody Formation; D022801:Complementarity Determining Regions / Q000235:genetics*; D019020:Directed Molecular Evolution / Q000379:methods*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D008832:Microchemistry / Q000379:methods*; D019151:Peptide Library; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D011995:Recombination, Genetic / Q000235:genetics
chemical_list: D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell