Soluble and stable heterodimeric TCR
Disclosed are a heterodimeric TCR containing artificial interchain disulfide bond between the variable region of α chain and the constant region of β chain, a preparing method therefor and a use thereof.
1. A αβ heterodimeric TCR, wherein an artificial interchain disulfide bond is contained between α chain variable region and β chain constant region of the TCR; and
cysteine residues that form the artificial interchain disulfide bond of the TCR substitute for:
an amino acid residue at position 46 of TRAV and an amino acid residue at position 60 of TRBC1*01 or TRBC2*01 exon 1;
an amino acid residue at position 47 of TRAV and an amino acid residue at position 61 of TRBC1*01 or TRBC2*01 exon 1;
an amino acid residue at position 46 of TRAV and an amino acid residue at position 61 of TRBC1*01 or TRBC2*01 exon 1; or
an amino acid residue at position 47 of TRAV and an amino acid residue at position 60 of TRBC1*01 or TRBC2*01 exon 1.
2. The TCR of claim 1 , wherein the TCR is soluble.
3. The TCR of claim 1 , wherein the TCR comprises α chain variable domain and β chain variable domain as well as all or part of β chain constant domains other than its transmembrane domain, however it does not comprise α chain constant domain, and α chain variable domain and β chain of the TCR form a heterodimer.
4. The TCR of claim 3 , wherein the cysteine residue in β chain constant domain for forming a natural interchain disulfide bond is replaced with another amino acid.
5. The TCR of claim 4 , wherein the cysteine residue in β chain constant domain for forming a natural interchain disulfide bond is replaced with alanine or serine.
6. The TCR of claim 3 , wherein the β chain constant domain of the TCR is truncated at C-terminus, thereby removing cysteine residues for forming natural interchain disulfide bonds.
7. The TCR of claim 1 , wherein the TCR comprises: (i) all or part of the TCR α chain other than its transmembrane domain, and (ii) all or part of the TCR β chain other than its transmembrane domain, wherein both of (i) and (ii) comprise variable domain and at least a portion of constant domains of TCR chain.
8. The TCR of claim 7 , wherein there is no natural interchain disulfide bond between α and β chain constant domain of the TCR.
9. The TCR of claim 8 , wherein the α chain and/or β chain constant region of the TCR are truncated at C-terminus, thereby removing cysteine residues for forming natural interchain disulfide bonds.
10. The TCR of claim 8 , wherein the cysteine residue in α chain and/or β chain constant region of the TCR for forming a natural interchain disulfide bond is substituted with another residue.
11. The TCR of claim 7 , wherein there is an artificial interchain disulfide bond between α chain constant region and β chain constant region of the TCR.
12. The TCR of claim 11 , wherein cysteine residues that form the artificial interchain disulfide bond between α chain constant region and β chain constant region of the TCR substitute for:
48T of TRAC1*01 exon 1 and 57S of TRBC1*01 or TRBC2*01 exon 1;
45T of TRAC1*01 exon 1 and 77S of TRBC1*01 or TRBC2*01 exon 1;
10Y of TRAC1*01 exon 1 and 17S of TRBC1*01 or TRBC2*01 exon 1;
45T of TRAC1*01 exon 1 and 59D of TRBC1*01 or TRBC2*01 exon 1;
15S of TRAC1*01 exon 1 and 15E of TRBC1*01 or TRBC2*01 exon 1;
53R of TRAC1*01 exon 1 and 54S of TRBC1*01 or TRBC2*01 exon 1;
89P of TRAC1*01 exon 1 and 19A of TRBC1*01 or TRBC2*01 exon 1; or
10Y of TRAC1*01 exon 1 and 20E of TRBC1*01 or TRBC2*01 exon 1.
13. The TCR of claim 1 , wherein a conjugate is bound with C- or N-terminus of the TCR α chain and/or β chain.
14. The TCR of claim 13 , wherein the conjugate bound with the TCR is selected from a group consisting of: a detectable marker; a therapeutic agent; a PK modifying moiety and a combination thereof.
15. The TCR of claim 14 , wherein the therapeutic agent bound with the TCR is anti-CD3 antibody which is linked at C- or N-terminus of α and/or β chains of the TCR.
16. A T-cell receptor complex, comprising one or more TCR molecules of claim 1 .
17. A pharmaceutical composition, comprising a pharmaceutically acceptable carrier and a safe and effective dosage of the TCR of claim 1 .