Method for improving the specific effector function of single-chain antigen-recognizing genetic constructs (scARC) through murinization thereof

摘要

The present invention relates to a method for producing a cell line expressing a stabilized functional single chain-antigen-recognizing genetic construct (scARC), comprising a genetic construct of the human scARC to be expressed, comprising the domains huV1/2—Li-huV2/1—C4/3 and a genetic construct comprising the corresponding hetero-/(homo-)dimeric domain C3/4, containing xenogenic, in its special case murine amino acid exchanges in the domains C4/3 and C3/4, wherein co-expression of the genetic constructs of the scARC-fragments occurs through the cell. Preferably, the scARCs are single chain-TCRs (scTCRs) or antibody-scFv-fragments, which further preferably recognize tumor associated peptide antigens (TAA). The present invention further relates to a gp100-protein-specific T-cell response mediated α/β T-cell receptor rationally mutated by means of the method of the present invention and its uses.

主权项

1. A method for producing an scARC expressing cell, the method comprising:
a) providing a suitable cell, b) providing a first genetic construct comprising the domains V1/2-Li-V2/1-C4/3, V1-Li-V2-C4 or V2-Li-V1-C3 and providing a second genetic construct comprising the corresponding hetero-/(homo-)dimeric domain C3/4, C3/4, containing xenogenic amino acid exchanges in the domains C4/3 and C3/4, and wherein the linker Li represents an arbitrary peptide sequence, c) introducing directly or indirectly the first and the second genetic constructs via viral or non-viral gene transfer into the cell, and d) co-expressing the first and the second genetic constructs in the cell, wherein the scARC is a single chain T-cell receptor (scTCR) and wherein only the ecto-subdomain of the constant domains contain the xenogenic amino acid exchanges.