Anti-CD33 antibody agents
Provided are anti-CD33 antibody agents, including anti-CD33 antibody agents in an IgG-scFv format. Various methods and reagents related thereto are also provided, including for example for the detection, prevention, and/or therapeutical treatment of CD33-related diseases, in particular, leukemias such as AML.
1. A bispecific antibody agent comprising two identical immunoglobulin heavy chains and two identical fusion polypeptides, wherein each heavy chain comprises a heavy chain variable domain sequence that is SEQ ID NO:1 or 2, and wherein each fusion polypeptide comprises:
(i) an immunoglobulin light chain comprising a light chain variable domain sequence that is SEQ ID NO:3 or 4, and
(ii) a single chain variable fragment (scFv),
wherein the heavy chain and the fusion polypeptide light chain bind to CD33, and wherein the fusion polypeptide scFv binds to CD3.
2. The bispecific antibody agent of claim 1 , wherein:
(i) the heavy chains each comprise a sequence identified as any one of SEQ ID NO:9, 20-22 and 24; and/or
(ii) the fusion polypeptide light chain comprises a light chain sequence identified as SEQ ID NO:10; and/or
(iii) the scFv is fused to the C-terminus of the immunoglobulin light chain.
3. The bispecific antibody agent of claim 1 , wherein the fusion polypeptide further comprises a linker.
4. The bispecific antibody agent of claim 1 , wherein:
(i) the fusion polypeptide comprises an scFv comprising a sequence identified as any of SEQ ID NOs: 11-17 and 27; or
(ii) the fusion polypeptides each comprise a sequence identified as SEQ ID NO: 26.
5. The bispecific antibody agent of claim 1 , wherein:
(i) the heavy chain comprising an Fc region with one or more mutations selected from K322A and D265A; or
(ii) the heavy chains each comprise a sequence identified as SEQ ID NO: 24.
6. An isolated nucleic acid molecule encoding a bispecific antibody agent of claim 1 .
7. A host cell comprising the isolated nucleic acid molecule of claim 6 .
8. A method for the production of a bispecific antibody agent comprising a step of culturing the host cell according to claim 7 in a culture medium under conditions allowing the expression of the antibody or fragment thereof and separating the antibody or fragment thereof from the culture medium.
9. A composition comprising the bispecific antibody agent of claim 1 .
10. A pharmaceutical composition comprising the composition of claim 9 , and further comprising a pharmaceutically acceptable carrier or diluent.
11. A T cell or a population of T cells armed with the bispecific binding agent of claim 1 .
12. A chimeric antigen receptor (CAR) comprising a bispecific binding agent of claim 1 .
13. A CAR-T cell, a population of CAR-T cells, a CAR-NK cell, or a population of CAR-NK cells expressing a CAR of claim 12 .
14. A composition comprising the population of CAR-NK cells of claim 13 .
15. The bispecific antibody agent of claim 1 , wherein the fusion polypeptide comprises an scFv that is a humanized OKT3 scFv.
16. The bispecific antibody agent of claim 1 , wherein the heavy chain variable domain comprises a sequence of SEQ ID NO:1 or 2, and wherein each fusion polypeptide comprises:
(i) an immunoglobulin light chain comprising a light chain variable domain sequence of SEQ ID NO:3 or 4, and
(ii) a single chain variable fragment (scFv) comprising a sequence of any one of SEQ ID NOs: 11-17 and 27.
17. The bispecific antibody agent of claim 1 , wherein:
(i) the heavy chains each comprise a sequence identified as SEQ ID NO: 24; and
(ii) the fusion polypeptides each comprise a sequence identified as SEQ ID NO: 26.
18. The isolated nucleic acid molecule of claim 6 , wherein the isolated nucleic acid comprises:
(i) a sequence encoding a heavy chain identified as SEQ ID NO: 23; and/or
(ii) a sequence encoding a fusion polypeptide identified as SEQ ID NO: 25.
19. A recombinant vector encoding the bispecific antibody agent of claim 1 .