soluable DMF5 TcR with TcRa_flag and TcRb-BirA
Biotinylation of the BirA sequence containing sTCR proteins
sTCRs recognize intracellular in addition to extracellular epitopes, potentially increasing the number of applications as reagents for target detection and immunotherapy. recently it has been show that they can be used for identification of their natural peptide ligands in disease.
Human cells were used as the platform of production and delivery of sTCRs because a) production in mammalian cells allows post-translational modifications; b) it permits the use of bicistronic vectors encoding both the TCRα and β chains separated by the ribosomal skipping sequence 2A found in the picorna virus
These produced sTCRs can be used for antigen-specific labeling and elimination of human target cells. Different TCRs were solubilized by expression of constructs encoding the TCR alpha (α) and beta (β) chains lacking the transmembrane and intracellular domains, linked by a ribosomal skipping 2A sequence that facilitates equimolar production of the chains. Cell supernatants containing sTCRs labeled target cells directly in a peptide (p)-human leukocyte antigen (HLA)-specific manner.