RECOMBINANT ANTIBODIES THAT RECONGNIZE THE C-TERMINAL DOMAINS OF EBOLA VIRUS NUCLEOPROTEIN

• US 20180334494A1
• Filed: 10/16/2015
• Published: 11/22/2018
• Est. Priority Date: 10/17/2014
• Status: Active Grant

• Current Assignee: University of Chicago, University of Virginia Patent Foundation (University of Virginia)
• Original Assignee: University of Chicago, University of Virginia Patent Foundation (University of Virginia)
• Inventors: DEREWENDA, Zygmunt S., ENGEL, Daniel A., KOSSIAKOFF, Anthony, DAVYDOVA, Elena K., JASKOLOWSKI, Mateusz
• Granted Patent: US 10,640,549 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 39/42 viral
• CPC Class Codes: C07K 16/10 from RNA viruses
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 2760/14111 Ebolavirus, e.g. Zaire ebolavirus
• CPC Class Codes: C12N 2760/14122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12Q 1/701 Specific hybridization probes
• CPC Class Codes: G01N 2333/08 RNA viruses
• CPC Class Codes: G01N 33/56983 Viruses

Abstract

This disclosure is directed to compositions and methods for utilizing the boundaries of the C-terminal domains of Nucleoprotein from Zaire Ebola virus as highly stable recombinant protein antigens to generate antibodies for diagnosis and treatment of Ebola virus infection.

Claims

1. An antibody that specifically binds to the Nucleoprotein of Ebola virus, or a binding fragment thereof, wherein said antibody comprises complementarity determining region (CDR) sequences selected from one of the following sets:

a) SEQ ID NO;

57, SEQ ID NO;

34, SEQ ID NO;

121, and SEQ ID NO;

35;

b) SEQ ID NO;

127, SEQ ID NO;

128, SEQ ID NO;

126, and SEQ ID NO;

129;

c) SEQ ID NO;

63, SEQ ID NO;

131, SEQ ID NO;

130, and SEQ ID NO;

132;

d) SEQ ID NO;

43, SEQ ID NO;

96, SEQ ID NO;

145, and SEQ ID NO;

35;

e) SEQ ID NO;

95, SEQ ID NO;

96, SEQ ID NO;

94, and SEQ ID NO;

97; and

f) SEQ ID NO;

105, SEQ ID NO;

51, SEQ ID NO;

104, and SEQ ID NO;

106.

2. The antibody of claim 1 wherein said antibody comprises complementarity determining region (CDR) sequences selected from one of the following sets:

a) SEQ ID NO;

105, SEQ ID NO;

51, SEQ ID NO;

104, and SEQ ID NO;

106; and

b) SEQ ID NO;

43, SEQ ID NO;

96, SEQ ID NO;

145, and SEQ ID NO;

35.

3. The antibody of claim 1, wherein the antibody is a monoclonal antibody.

4. The antibody of claim 1, wherein the antibody is a chimeric antibody.

5. The antibody of claim 1, wherein the antibody is a fragment binding protein (Fab).

7. The antibody of claim 1, wherein the antibody is expressed as a recombinant antibody having a variant Fc domain.

8. The antibody of claim 1, wherein said antibody binds the C-terminal domain of the Nucleoprotein of an Ebola virus strain selected from the group consisting of Tai Forest, Sudan, Bundibugyo, Reston, and Zaire.

9. The antibody of claim 8, wherein said antibody binds an amino acid sequence selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, and SEQ ID NO;

5.

10. The antibody of claim 1, wherein the antibody comprises a heavy chain sequence of SEQ ID NO:

210 and a light chain of SEQ ID NO;

211 or comprises a heavy chain of SEQ ID NO;

212 and a light chain of SEQ ID NO;

213.

11. The antibody of claim 10, wherein the antibody is a Fab fragment consisting of a heavy chain sequence of SEQ ID NO:

210 and a light chain of SEQ ID NO;

211.

12. The antibody of claim 10, wherein the antibody is a Fab fragment consisting of a heavy chain sequence of SEQ ID NO:

212 and a light chain of SEQ ID NO;

213.

17. A diagnostic kit comprising the antibody of claim 1 and reagents for detecting the binding of the antibody to its target antigen.

18. A pharmaceutical composition comprising the antibody of claim 1, and a pharmaceutically acceptable carrier, excipient, or diluent.

19. A method of screening a biological sample to detect the presence of an Ebola viral protein, the method comprising

contacting the biological sample with the antibody of claim 1;

and

detecting the specific binding of said antibody with a Nucleoprotein in the biological sample.

20. The method of claim 19 wherein the antibody is immobilized on a substrate and detection is based on the binding of a labeled secondary antibody to said Nucleoprotein.

21. The method of claim 20 wherein the immobilized antibody and the secondary antibody are selected from the group consisting of Fab6 and Fab20.

22. A test strip for detecting Ebola virus, said test strip comprising

a solid support;

and

an antibody of claim 1 immobilized on the surface of the solid support.

6. (canceled)

13. An antibody that specifically binds to the Nucleoprotein of Ebola virus, or a binding fragment thereof, wherein the complementarity determining regions comprise one or more amino acid sequences selected from the group consisting of SEQ ID NO:

105, SEQ ID NO;

51, SEQ ID NO;

104, SEQ ID NO;

106, SEQ ID NO;

35, SEQ ID NO;

145, SEQ ID NO;

96, and SEQ ID NO;

43.

14. The antibody of claim 13, wherein the antibody is labeled with a detectable moiety.

15. An antibody that binds to a recombinantly expressed Ebola virus Nucleoprotein domain selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4 and SEQ ID NO;

5, wherein the CDRs of said antibody bind to noncontiguous regions of said protein domain.

16. The antibody of claim 15, wherein the antibody is isolated using a phage display library.