Directed evolution of human T-cell receptors with picomolar affinities by phage display.
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.
Li
Yi
Y
Moysey
Ruth
R
Molloy
Peter E
PE
Vuidepot
Anne-Lise
AL
Mahon
Tara
T
Baston
Emma
E
Dunn
Steven
S
Liddy
Nathaniel
N
Jacob
Jansen
J
Jakobsen
Bent K
BK
Boulter
Jonathan M
JM
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
D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell