Patent ID: 9062127
Filing Date: 2015-06-23
Classification: A61P,C07H,C07K,C12N

Abstract:
1. An in vitro method for producing a heterodimeric specific wild type- or chimeric T-cell receptor (TCR), said TCR being a human TCR, a murine TCR or a humanized murine TCR, the method comprising: (a) providing DNA-molecules in joint or separate mutagenesis vector system(s), the DNA-molecules comprising the coding regions for a first and second chain of a TCR, said first and second chain of said TCR interacting at at least one surface in the constant regions of the first and second chains, (b) mutagenizing the DNA-molecules to produce individual mutated DNA-molecules, wherein the nucleic acid sequence encoding the first chain is mutagenized to encode the first chain comprising a substitution in an amino acid in the at least one surface within the constant region of the first chain, wherein the amino acid interacts with the second chain and the amino acid introduced after mutagenesis in the at least one surface of the first chain provides a sterically projecting group in the constant domain of the first chain and the nucleic acid sequence encoding the second chain is mutagenized to encode the second chain comprising a substitution in an amino acid in the at least one surface within the constant region that of the second chain, wherein the amino acid interacts with the first chain and the amino acid introduced after mutagenesis in the at least one surface of the second chain provides a sterically recessed group in the constant domain of the second chain, (c) translating the mutated DNA molecules from step (b), such that pairing of the heterodimeric specific first-chain/second-chain TCR being mutated at the at least one surface within the constant region is selectively promoted, and (d) expressing in vitro the heterodimeric first-chain/second-chain TCR by a T-cell, wherein the first chain and second chain of said TCR interact via the sterically projecting group and the sterically recessed group at said at least one surface located in the constant regions of said first and second chains wherein an amino acid that has been introduced after the mutagenesis of the DNA-molecules that introduces a sterically recessing group compared to the initial sequence is selected from glycine, serine, threonine, valine and alanine, and wherein an amino acid that has been introduced after the mutagenesis of the DNA-molecules that introduces a sterically projecting group compared to the initial sequence is selected from tryptophan, lysine, arginine, phenylalanine, cysteine and tyrosine.