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

Claim 1:
2. A method for the identification of a TCR recognizable peptide sequence, comprising:
i. providing a mammalian cell, wherein
said mammalian cell expresses a transgenic antigen receptor;
said transgenic antigen receptor molecule comprises an oligopeptide 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 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 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; and
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 mammalian cell with a preparation of T-lymphocytes, thereby activating said reporter gene yielding an activated mammalian cell,
iii. isolating DNA from said activated mammalian cell, and
iv. sequencing of said oligopeptide sequence encoded in said transgenic chimeric antigen-receptor.