Modified soluble T cell receptor
View Patent ↗The present invention provides a soluble T cell receptor (sTCR), which comprises (i) all or part of a TCR $g(a) chain, except the transmembrane domain thereof, and (ii) all or part of a TCR $g(b) chain, except the transmembrane domain thereof. (i) and (ii) each comprise a functional variable domain and at least a part of the constant domain of the TCR chain, and are linked by a disulphide bond between constant domain residues which is not present in native TCR, characterized in that the sTCR recognizes a CD1-antigen complex, a bacterial superantigen or a peptide-MHC/superantigen complex.
1. A soluble T cell receptor (sTCR), which comprises (i) all or part of a TCR α chain, except the transmembrane domain thereof, and (ii) all or part of a TCR β chain, except the transmembrane domain thereof, wherein (i) and (ii) each comprise a variable domain and at least a part of the constant domain of the TCR chain, and are linked by a disulphide bond between constant domain cysteine residues substituted for Thr 48 of exon 1 of TRAC*01 and Ser 57 of exon 1 of TRBC1*01 or TRBC2*01, wherein Thr 48 is amino acid 10 of SEQ ID NO:1 and wherein Ser 57 is amino acid 10 of SEQ ID NO:5 characterized in that the sTCR binds a CD1-antigen complex, wherein one or both of (i) and (ii) comprise all of the extracellular constant immunoglobulin domain of the TCR chain.
2. A sTCR as claimed in claim 1 , wherein one or both of (i) and (ii) comprise all of the extracellular domain of the TCR chain.
3. A sTCR as claimed in claim 1 , wherein an interchain disulphide bond in native TCR is not present.
4. A sTCR as claimed in claim 3 , wherein native α and β TCR chains are truncated at the C-terminus such that the cysteine residues which form the native interchain disulphide bond are excluded.
5. A sTCR as claimed in claim 3 , wherein cysteine residues which form the native interchain disulphide bond are substituted to another residue.
6. A sTCR as claimed in claim 5 , wherein cysteine residues which form the native interchain disulphide bond are substituted to serine or alanine.
7. A sTCR as claimed in claim 1 , wherein an unpaired cysteine residue present in native TCR β chain is not present.
8. A sTCR as claimed in claim 1 , wherein (i) and (ii) each comprise the variable domain of a first TCR fused to all or part of the constant domain of a second TCR, the first and second TCRs being from the same species.
9. A sTCR as claimed in claim 1 , wherein one or both of the chains are fused to a moiety at its C or N terminus.
10. A sTCR as claimed in claim 1 , wherein one or both of the chains have a cysteine residue at its C and/or N terminus to which a moiety can be fused.
11. A sTCR as claimed in claim 1 , further comprising a detectable label.
12. A sTCR as claimed in claim 1 associated with a therapeutic agent.
13. A multivalent T cell receptor (TCR) complex comprising a plurality of sTCRs as claimed in claim 1 .
14. A complex as claimed in claim 13 , comprising a sTCR multimer.
15. A complex as claimed in claim 14 , wherein the sTCR multimer comprises two or three or four or more sTCRs associated with one another.
16. A complex as claimed in claim 13 , wherein the sTCRs or sTCR multimers are present in a lipid bilayer or are attached to a particle.
17. A method for detecting CD1-antigen complexes, which comprises:
(i) providing a soluble TCR as claimed in claim 1 or a multivalent T cell receptor complex comprising a plurality of sTCRs as claimed in claim 1 ;
(ii) contacting the soluble TCR or multivalent TCR complex with the CD-1antigen complexes; and
(iii) detecting binding of the soluble TCR or multivalent TCR complex to the CD-1 antigen complexes.
18. A pharmaceutical formulation comprising a sTCR as claimed in claim 1 , and/or a multivalent TCR complex comprising a plurality of sTCRs as claimed in claim 1 , together with a pharmaceutically acceptable carrier.
19. The complex of claim 15 wherein the sTCRs are associated with one another via a linker molecule.