Soluble T cell receptor
View Patent ↗The present invention provides a soluble T cell receptor (sTCR), which comprises (i) all for 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. (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.
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 human TCR β chain, except the transmembrane domain thereof, wherein (i) and (ii) each comprise a functional variable domain and at least a part of the constant domain of the TCR chain, characterised in that (i) and (ii) are linked by a disulphide bond between cysteine residues substituted for:
Thr 48 of exon 1 of TRAC*01 (amino acid residue 10 of SEQ ID NO:1) and Ser 57 of exon 1 of TRBC1*01 or TRBC2*01 (amino acid residue 10 of SEQ ID NO:5);
Thr 45 of exon 1 of TRAC*01 (amino acid residue 10 of SEQ ID NO:2) and Ser 77 of exon 1 of TRBC1*01 or TRBC2*01 (amino acid residue 10 of SEQ ID NO:6);
Tyr 10 of exon 1 of TRAC*01 (amino acid residue 10 of SEQ ID NO:3) and Ser 17 of exon 1 of TRBC1*01 or TRBC2*01 (amino acid residue 10 of SEQ ID NO:7);
Thr 45 of exon 1 of TRAC*01 (amino acid residue 10 of SEQ ID NO:2) and Asp 59 of exon 1 of TRBC1*01 or TRBC2*01 (amino acid residue 10 of SEQ ID NO:8); or
Ser 15 of exon 1 of TRAC*01 (amino acid residue 10 of SEQ ID NO:4) and Glu 15 of exon 1 of TRBC1*01 or TRBC2*01 (amino acid residue 10 of SEQ ID NO:9).
2. The sTCR as claimed in claim 1 , wherein one or both of (i) and (ii) comprise all of the extracellular constant Ig domain of the TCR chain.
3. The sTCR as claimed in claim 1 , wherein one or both of (i) and (ii) comprise all of the extracellular domain of the TCR chain.
4. A soluble αβ-form T cell receptor (sTCR), wherein a covalent disulphide bond links cysteine residues substituted for:
Thr 48 of exon 1 of TRAC*01 (amino acid residue 10 of SEQ ID NO:1)and Ser 57 of exon 1 of TRBC1*01 or TRBC2*01 (amino acid residue 10 of SEQ ID NO:5).
5. The sTCR as claimed in claim 1 , wherein an interchain disulphide bond in native TCR is not present.
6. The sTCR as claimed in claim 5 , 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.
7. The sTCR as claimed in claim 5 , wherein cysteine residues which form the native interchain disulphide bond are substituted to another residue.
8. The sTCR as claimed in claim 7 , wherein cysteine residues which form the native interchain disulphide bond are substituted to serine or alanine.
9. The sTCR as claimed in claim 1 , wherein an unpaired cysteine residue present in native TCR β chain is not present.
10. The sTCR as claimed in any one of claims 1 , wherein (i) and (ii) each comprise the functional variable domain of a first human TCR fused to all or part of the constant domain of a human second TCR.
11. The sTCR as claimed in claim 1 , wherein one or both of the chains are fused to a moiety at its C or N terminus.
12. The 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.
13. The sTCR as claimed claim 1 , further comprising a detectable label.
14. The sTCR as claimed in claim 1 associated with a therapeutic agent.
15. A multivalent T cell receptor (TCR) complex comprising a plurality of sTCRs as claimed in claim 1 .
16. The complex as claimed in claim 15 , comprising a sTCR multimer.
17. The complex as claimed in claim 16 , comprising two or three or four or more T cell receptor molecules associated with one another, preferably via a linker molecule.
18. The complex as claimed in claim 15 , wherein the sTCRs or sTCR multimers are present in a lipid bilayer or are attached to a particle.
19. A method for detecting MHC-peptide complexes, which comprises:
(i) providing the soluble TCR as claimed in claim 1 ;
(ii) contacting the soluble TCR or with the MHC-peptide complexes; and
(iii) detecting binding of the soluble TCR to the MHC-peptide complexes.
20. A pharmaceutical formulation comprising the sTCR as claimed in claim 1 together with a pharmaceutically acceptable carrier.
21. A pharmaceutical formulation comprising the multivalent TCR complex as claimed in claim 15 together with a pharmaceutically acceptable carrier.
22. A method for detecting MHC-peptide complexes, which comprises:
(i) providing the multivalent T cell receptor complex as claimed in claim 15 ;
(ii) contacting the multivalent TCR complex with the MHC-peptide complexes; and
(iii) detecting binding of the multivalent TCR complex to the MHC-peptide complexes.