CD33-binding polypeptides and uses thereof
Provided herein are VHH-containing polypeptides that bind CD33. Uses of the VHH-containing polypeptides are also provided.
1. An isolated nucleic acid that encodes a polypeptide comprising at least one VHH domain that binds CD33, wherein at least one VHH domain that binds CD33 comprises:
a) a CDR1 comprising the amino acid sequence of SEQ ID NO: 47; a CDR2 comprising the amino acid sequence of SEQ ID NO: 48; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 49;
b) a CDR1 comprising the amino acid sequence of SEQ ID NO: 7; a CDR2 comprising the amino acid sequence of SEQ ID NO: 8; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 9;
c) a CDR1 comprising the amino acid sequence of SEQ ID NO: 50; a CDR2 comprising the amino acid sequence of SEQ ID NO: 51; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 52;
d) a CDR1 comprising the amino acid sequence of SEQ ID NO: 56; a CDR2 comprising the amino acid sequence of SEQ ID NO: 54 or 57; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 58;
e) a CDR1 comprising the amino acid sequence of SEQ ID NO: 59; a CDR2 comprising the amino acid sequence of SEQ ID NO: 60; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 61;
f) a CDR1 comprising the amino acid sequence of SEQ ID NO: 23 or 62; a CDR2 comprising the amino acid sequence of SEQ ID NO: 63; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 64;
g) a CDR1 comprising the amino acid sequence of SEQ ID NO: 65; a CDR2 comprising the amino acid sequence of SEQ ID NO: 66; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 67;
h) a CDR1 comprising the amino acid sequence of SEQ ID NO: 68; a CDR2 comprising the amino acid sequence of SEQ ID NO: 69; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 70; or
i) a CDR1 comprising the amino acid sequence of SEQ ID NO: 71; a CDR2 comprising the amino acid sequence of SEQ ID NO: 72; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 73.
2. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 47; a CDR2 comprising the amino acid sequence of SEQ ID NO: 48; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 49.
3. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 7; a CDR2 comprising the amino acid sequence of SEQ ID NO: 8; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 9.
4. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 50; a CDR2 comprising the amino acid sequence of SEQ ID NO: 51; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 52.
5. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 56; a CDR2 comprising the amino acid sequence of SEQ ID NO: 54 or 57; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 58.
6. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 59; a CDR2 comprising the amino acid sequence of SEQ ID NO: 60; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 61.
7. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 23 or 62; a CDR2 comprising the amino acid sequence of SEQ ID NO: 63; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 64.
8. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 65; a CDR2 comprising the amino acid sequence of SEQ ID NO: 66; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 67.
9. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 68; a CDR2 comprising the amino acid sequence of SEQ ID NO: 69; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 70.
10. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 71; a CDR2 comprising the amino acid sequence of SEQ ID NO: 72; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 73.
11. The isolated nucleic acid of claim 1 , wherein at least one VHH domain is humanized.
12. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 38, 114, 39, 40, 41, 42, 43, 44, 45, 46, 115, 116, 117, 118, 119, 120, 121, or 122.
13. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38, 114, 39, 40, 41, 42, 43, 44, 45, 46, 115, 116, 117, 118, 119, 120, 121, or 122.
14. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38 or 114.
15. The isolated nucleic acid of claim 1 , wherein at least one VHH domain comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 123, 124, 125, 126, 127, 128, 129, 130, or 131.
16. The isolated nucleic acid of claim 1 , wherein the polypeptide comprises one, two, or three VHH domains.
17. The isolated nucleic acid of claim 1 , wherein the polypeptide comprises at least one binding domain that binds an antigen other than CD33.
18. The nucleic acid of claim 17 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38 or 114.
19. The isolated nucleic acid of claim 17 , wherein the polypeptide comprises at least one binding domain that binds CD3, T-cell receptor (TCR) α, TCRβ, CD28, CD16, CD32A, CD64, CD89, NKp46, or NKG2D.
20. The isolated nucleic acid of claim 16 , wherein each VHH domain binds CD33.
21. The nucleic acid of claim 20 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38 or 114.
22. The isolated nucleic acid of claim 1 , wherein the polypeptide comprises an Fc region.
23. The isolated nucleic acid of claim 22 , wherein the Fc region comprises an amino acid sequence selected from SEQ ID NOs: 74-109.
24. A vector comprising the isolated nucleic acid of claim 1 .
25. A host cell comprising the isolated nucleic acid of claim 1 .
26. The host cell of claim 25 , wherein at least one VHH domain comprises the amino acid sequence of SEQ ID NO: 38 or 114.
27. A method of producing a polypeptide, comprising incubating the host cell of claim 25 under conditions suitable for expression of the polypeptide, and isolating the polypeptide.