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US 9,751,928 B2Granted
Enhanced affinity T cell receptors and methods for making the same
AssigneeFred Hutchinson Cancer Research Center
InventorsThomas M. Schmitt, Philip D. Greenberg
Life of this patent
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Abstract
The present disclosure provides methods for generating enhanced affinity T cell receptors by agonist selection of hematopoietic progenitor cells expressing an antigen specific TCRα cultured with stromal cells expressing Delta-like-1 or Delta-like-4, compositions prepared from such methods, and uses of thereof.
Claims20
A method for generating an enhanced affinity T cell receptor (TCR), comprising:
- a. contacting one or more hematopoietic progenitor cells with one or more stromal cells and a peptide antigen, under conditions and for a time sufficient to induce differentiation of at least one of the hematopoietic progenitor cells into a double negative (DN) TCRαβ30 thymocyte,
- wherein the one or more hematopoietic progenitor cells comprise a non-endogenous polynucleotide encoding a TCRα chain from a parent TCR specific for the peptide antigen, and
- wherein the one or more stromal cells comprise a non-endogenous polynucleotide encoding Delta-like-1 or Delta-like-4 and a polynucleotide encoding an MHC molecule;
- b. introducing at least one polynucleotide encoding a TCRβ chain isolated from the DN TCRαβ+thymocyte generated in step (a) into one or more cells that are capable of expressing a TCR on the cell surface and comprise the non-endogenous polynucleotide encoding the TCRα chain from step (a), thereby generating one or more cells expressing a candidate TCR;
- and
- c. identifying an enhanced affinity TCR expressed by at least one of the one or more cells generated in step (b),
- wherein identifying the enhanced affinity TCR comprises comparing the binding affinity of the candidate TCR to the binding affinity of the parent TCR.
4 more elements
Dependencies are inferred from the claim text.