Single-chain chimeric polypeptides and uses thereof
Provided herein are single-chain chimeric polypeptides that include: (i) a first target-binding domain; (ii) a soluble tissue factor domain; and (iii) a second target-binding domain. Also provided here are methods of using these single-chain chimeric polypeptides and nucleic acids encoding these single-chain chimeric polypeptides.
1. A nucleic acid encoding a single-chain chimeric polypeptide comprising:
(i) a first target-binding domain comprising a sequence at least 80% identical to SEQ ID NO: 28;
(ii) a soluble tissue factor domain that does not have coagulation activity, wherein the soluble tissue factor domain comprises a sequence at least 80% identical to SEQ ID NO: 9; and
(iii) a second target-binding domain comprising a sequence at least 80% identical to SEQ ID NO: 28,
wherein the single-chain chimeric polypeptide does not stimulate blood coagulation in a mammal.
2. The nucleic acid of claim 1 , wherein the first target-binding domain and the soluble tissue factor domain directly abut each other.
3. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the soluble tissue factor domain.
4. The nucleic acid of claim 1 , wherein soluble tissue factor domain and the second target-binding domain directly abut each other.
5. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide further comprises a linker sequence between the soluble tissue factor domain and the second target-binding domain.
6. The nucleic acid of claim 1 , wherein the soluble tissue factor domain is a soluble human tissue factor domain that does not have coagulation activity.
7. The nucleic acid of claim 1 , wherein the soluble tissue factor domain comprises or consists of a sequence from a wildtype soluble human tissue factor.
8. The nucleic acid of claim 1 , wherein the first target-binding domain and the second target-binding domain are human IL-2.
9. A vector comprising the nucleic acid of claim 1 .
10. A cell comprising the nucleic acid of claim 1 .
11. A method of producing a single-chain chimeric polypeptide, the method comprising:
culturing the cell of claim 10 in a culture medium under conditions sufficient to result in the production of the single-chain chimeric polypeptide; and
recovering the single-chain chimeric polypeptide from the cell and/or the culture medium.
12. The nucleic acid of claim 1 , wherein:
the first target-binding domain comprises a sequence at least 90% identical to SEQ ID NO: 28;
the soluble tissue factor domain comprises a sequence at least 90% identical to SEQ ID NO: 9; and
the second target-binding domain comprises a sequence at least 90% identical to SEQ ID NO: 28.
13. The nucleic acid of claim 1 , wherein:
the first target-binding domain comprises a sequence at least 95% identical to SEQ ID NO: 28;
the soluble tissue factor domain comprises a sequence at least 95% identical to SEQ ID NO: 9; and
the second target-binding domain comprises a sequence at least 95% identical to SEQ ID NO: 28.
14. The nucleic acid of claim 1 , wherein:
the first target-binding domain comprises SEQ ID NO: 28;
the soluble tissue factor domain comprises SEQ ID NO: 9; and
the second target-binding domain comprises SEQ ID NO: 28.
15. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 108.
16. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 108.
17. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide comprises a sequence that is at least 95% identical to SEQ ID NO: 108.
18. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide comprises SEQ ID NO: 108.
19. The nucleic acid of claim 18 , wherein the nucleic acid comprises the sequence of SEQ ID NO: 109.
20. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 110.
21. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 110.
22. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide comprises a sequence that is at least 95% identical to SEQ ID NO: 110.
23. The nucleic acid of claim 1 , wherein the single-chain chimeric polypeptide comprises SEQ ID NO: 110.
24. The nucleic acid of claim 22 , wherein the nucleic acid comprises the sequence of SEQ ID NO: 111.
25. A composition comprising the nucleic acid of claim 1 .
26. A kit comprising the nucleic acid of claim 1 .
27. The vector of claim 9 , wherein the vector is an expression vector.