Anti-CD20 antibodies and fusion proteins thereof and methods of use
The present invention provides humanized, chimeric and human anti-CD20 antibodies and CD 20 antibody fusion proteins that bind to a human B cell marker, referred to as CD20, which is useful for the treatment and diagnosis of B-cell disorders, such as B-cell malignancies and autoimmune diseases, and methods of treatment and diagnosis.
1. A humanized antibody or antigen-binding antibody fragment thereof that binds CD20 comprising hA20Vk (residues 20–125 of SEQ ID NO: 46) and hA20VH1 (SEQ ID NO: 41).
2. A humanized antibody or antigen-binding antibody fragment thereof that binds CD20 comprising hA20Vk (residues 20–125 of SEQ ID NO: 46) and hA20VH2 (SEQ ID NO: 42).
3. The humanized antibody and antigen-binding fragments of claim 1 , wherein said antigen-binding fragments are selected from the group consisting of F(ab′) 2 , Fab′, Fab, Fv and sFv.
4. An isolated DNA sequence comprising a nucleic acid encoding hA20Vk (residues 20–125 of SEQ ID NO: 46) and hA20VH1 (SEQ ID NO: 41).
5. An expression vector comprising the DNA sequence of claim 4 .
6. An isolated host cell comprising the DNA sequence of claim 4 .
7. A method for expression of an antibody or antigen-binding fragment thereof comprising hA20Vk (residues 20–125 of SEQ ID NO: 46) and hA20VH1 (SEQ ID NO: 41), comprising:
(a) transfecting a mammalian cell with a DNA sequence comprising a nucleic acid that encodes hA20Vk (residues 20–125 of SEQ ID NO: 46) and hA20VH1 (SEQ ID NO: 41); and
(b) culturing said cell secreting said antibody or antigen-binding fragment thereof.
8. A method of treating a B-cell lymphoma or leukemia in a subject comprising administering to said subject a therapeutically effective amount of an antibody or an antigen-binding fragment thereof as claimed in claim 1 .
9. A method of treating an autoimmune disease in a subject, wherein said autoimmune disease is mediated by B-cells, comprising administering to said subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof as claimed in claim 1 .
10. The humanized antibody and antigen-binding fragments of claim 2 , wherein said antigen-binding fragments are selected from the group consisting of F(ab′) 2 , Fab′, Fab, Fv and sFv.
11. An isolated DNA sequence comprising a nucleic acid encoding hA20Vk (residues 20–125 of SEQ ID NO: 46) and hA20VH2 (SEQ ID NO: 42).
12. An expression vector comprising the DNA sequence of claim 11 .
13. An isolated host cell comprising the DNA sequence of claim 11 .
14. A method for expression of an antibody or antigen-binding fragment thereof comprising hA20Vk (residues 20–125 of SEQ ID NO: 46) and hA20VH2 (SEQ ID NO: 42), comprising:
(a) transfecting a mammalian cell with a DNA sequence comprising a nucleic acid that encodes hA20Vk (residues 20–125of SEQ ID NO: 46) and hA20VH2 (SEQ ID NO: 42); and
(b) culturing said cell secreting said antibody or antigen-binding fragment thereof.
15. A method of treating a B-cell lymphoma or leukemia in a subject comprising administering to said subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof as claimed in claim 2 .
16. A method of treating an autoimmune disease in a subject, wherein said autoimmune disease is mediated by B-cells, comprising administering to said subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof as claimed in claim 2 .