Patent ID: 11939572
Assignee: ETH ZüRICH
Field: Pharmaceuticals (Chemistry)
Classification: CPC C  A  G | IPC A  C  G

Claim 0:
1. A method for the identification of a TCR recognizable peptide sequence, comprising:
i. providing a plurality of mammalian cells, wherein
each of said plurality of mammalian cells expresses a member of a library;
each member of said library encodes a transgenic antigen receptor;
said transgenic antigen receptor comprises an oligopeptide different for each member of the library and a chimeric antigen-receptor polypeptide heterodimer comprising a first polypeptide and a second polypeptide, wherein:
a. said first polypeptide comprises an extracellular part of a major histocompatibility complex I (MHC class I) alpha chain, wherein the oligopeptide is comprised within said extracellular part of the MHC class I alpha chain and presented in a way suitable for the recognition by a T cell receptor, and wherein said extracellular part of the MHC alpha chain retains the ability to interact with the oligopeptide and with CD8, and said second polypeptide comprises a β2-microglobulin domain, or
b. said first polypeptide comprises an extracellular part of a major histocompatibility complex II (MHC class II) alpha chain and said second polypeptide comprises an extracellular part of a MHC class II beta chain, wherein the oligopeptide is comprised within said extracellular parts of the MHC class II alpha and beta chains and presented in a way suitable for the recognition by a T cell receptor, and wherein said extracellular parts of the MHC alpha and beta chains retain the ability to interact with the oligopeptide and CD4,

one of said first polypeptide and said second polypeptide further comprises a hinge region, a transmembrane domain and an intracellular domain or intracellular tail of a T cell receptor alpha chain and the other one of said first polypeptide and said second polypeptide comprises a hinge region, a transmembrane domain and an intracellular domain of a T cell receptor beta chain;
said transgenic antigen receptor is functionally linked to a reporter gene, whereby binding of a cognate T cell receptor to said transgenic antigen receptor results in the activation of a reporter protein encoded by said reporter gene,

ii. contacting said plurality of mammalian cells with a preparation of T-lymphocytes,
iii. separating cells showing a detectable reporter protein from said plurality of mammalian cells according to the detectable level of said reporter protein, yielding activated cells,
iv. isolating DNA from said activated cells, and
v. sequencing of said oligopeptide sequence comprised in said transgenic antigen-receptor.