Single-domain VHH antibodies directed to norovirus GI.1 and GII.4 and their use

Patent metadata

Field	Value
Publication number	US-10000556-B2
Application number	US-201414889774-A
Country	US
Kind code	B2
Filing date	May 9, 2014
Priority date	May 9, 2013
Publication date	Jun 19, 2018
Grant date	Jun 19, 2018

Abstract

Isolated V H H monoclonal antibodies are disclosed that specifically bind to a Norovirus polypeptide. In some embodiments, the Norovirus is a Genogroup I Norovirus or a Genogroup II Norovirus. In other embodiments, the Norovirus is Norwalk or MD2004 virus. In some embodiments, the monoclonal antibodies specifically bind VP1. Also disclosed are compositions including the disclosed antibodies, nucleic acids encoding these antibodies, expression vectors including the nucleic acids, and isolated host cells that express the nucleic acids. The antibodies and compositions disclosed herein can be used for detecting the presence of a Norovirus in a biological sample, or detecting a Norovirus infection. Also disclosed are methods of treating and/or preventing a NoV infection.

First claim

We claim:

1. An isolated V H H monoclonal antibody, or an antigen binding fragment thereof, comprising a heavy chain domain, wherein the heavy chain variable domain comprises a heavy chain complementarity determining region (HCDR)1, an HCDR2 and an HCDR3, and wherein the V H H monoclonal antibody specifically binds a Norovirus (NoV) polypeptide, and wherein
a) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 3, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 3, and the CDR3 comprises amino acids 96-109 of SEQ ID NO: 3;
b) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 2, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 2, and the CDR3 comprises amino acids 96-109 of SEQ ID NO: 2;
c) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 1, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 1, and the CDR3 comprises amino acids 96-109 of SEQ ID NO: 1;
d) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 4, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 4, and the a CDR3 comprises amino acids 96-109 of SEQ ID NO: 4;
e) the CDR1 comprising amino acids 26-33 of SEQ ID NO: 5, the CDR2 comprising amino acids 51-58 of SEQ ID NO: 5, and the CDR3 comprising amino acids 97-110 of SEQ ID NO: 5;
f) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 6, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 6, and the CDR5 comprises amino acids 97-111 of SEQ ID NO: 6;
g) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 7, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 7, and the CDR3 comprises amino acids 97-111 of SEQ ID NO: 7;
h) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 8, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 8, and the CDR3 comprises amino acids 97-111 of SEQ ID NO: 8;
i) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 9, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 9, and the CDR3 comprises amino acids 96-112 of SEQ ID NO: 9;
j) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 10, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 10, and the CDR3 comprises amino acids 96-112 of SEQ ID NO: 10;
k) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 11, the CDR2 comprises amino acids 51-57, and the CDR3 comprises amino acids 96-114 of SEQ ID NO: 11;
l) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 12, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 12, and the CDR3 comprises amino acids 96-112 of SEQ ID NO: 12;
m) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 13, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 13, and the CDR3 comprises amino acids 97-113 of SEQ ID NO: 13;
n) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 14, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 14, and the CDR3 comprises amino acids 97-113 of SEQ ID NO: 14;
o) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 15, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 15, and the CDR3 comprises amino acids 97-114 of SEQ ID NO: 15;
p) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 16, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 16, and the CDR3 comprises amino acids 96-107 of SEQ ID NO: 16;
q) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 17, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 17, and the CDR3 comprises amino acids 97-113 of SEQ ID NO: 17;
r) the CDR1 comprising amino acids 26-33 of SEQ ID NO: 18, the CDR2 comprising amino acids 51-58 of SEQ ID NO: 18, and the CDR3 comprising amino acids 97-114 of SEQ ID NO: 18;
s) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 19, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 19, and the CDR3 comprises amino acids 97-113 of SEQ ID NO: 19;
t) the CDR1 comprising amino acids 26-33 of SEQ ID NO: 20, the CDR2 comprises amino acids 51-57 of SEQ ID NO:20, and the CDR3 comprises amino acids 96-111 of SEQ ID NO: 20; or
u) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 21, the CDR2 comprises amino acids 51-57 of SEQ ID NO:21, and the CDR3 comprises amino acids 96-102 of SEQ ID NO: 21.

2. The isolated V H H monoclonal antibody of claim 1 , wherein the heavy chain variable domain comprises the amino acid sequence set for the as one of SEQ ID NOs: 1-21.

3. The isolated monoclonal antibody of claim 1 , wherein the antibody is an IgG.

4. The isolated monoclonal antibody of claim 1 , wherein the antibody is humanized or chimeric.

5. The isolated antigen binding fragment of the isolated monoclonal antibody of claim 1 .

6. The isolated monoclonal antibody or the antigen binding fragment of claim 1 , wherein the antibody or antigen binding fragment is labeled.

7. The isolated monoclonal antibody or antigen binding fragment of claim 6 , wherein the label is a fluorescent, an enzymatic, or a radioactive label.

8. A composition comprising an effective amount of the antibody of claim 1 , or the antigen binding fragment of the antibody, and a pharmaceutically acceptable carrier.

9. An isolated nucleic acid molecule encoding the monoclonal antibody or antigen binding fragment of claim 1 .

10. A method of detecting a Norovirus infection in a subject comprising:
contacting a biological sample from the subject with at least one isolated monoclonal antibody of claim 1 or the antigen binding fragment thereof; and
detecting antibody bound to the sample,
wherein the presence of antibody bound to the sample indicates that the subject has an Norovirus infection.

11. The method of claim 10 , wherein the isolated monoclonal antibody or the antigen binding fragment is directly labeled.

12. The method of claim 10 , further comprising:
contacting the sample with a second antibody that specifically binds the isolated human monoclonal antibody; and
detecting the binding of the second antibody,
wherein an increase in binding of the second antibody to the sample as compared to binding of the second antibody to a control sample detects the presence of an Norovirus infection the subject.

13. A method for inhibiting a Norovirus infection in a subject, comprising:
administering to the subject a therapeutically effective amount of an antibody of claim 1 , an antigen binding fragment thereof, or a nucleic acid encoding the antibody or antigen binding fragment,
thereby inhibiting the Norovirus infection.

14. The method of claim 13 , further comprising administering to the subject an anti-viral agent.

15. The method of claim 13 , further comprising measuring viral titer in a biological sample from the subject.

16. A composition comprising an effective amount of the nucleic acid molecule of claim 9 ; or a vector comprising the nucleic acid molecule; and a pharmaceutically acceptable carrier.

17. A method for inhibiting a Norovirus infection in a subject, comprising:
administering to the subject a therapeutically effective amount of an antibody of claim 1 , an antigen binding fragment thereof, or a nucleic acid encoding the antibody or antigen binding fragment,
thereby inhibiting the Norovirus infection.

18. The method of claim 17 , further comprising administering to the subject an anti-viral agent.

19. An isolated V H H monoclonal antibody, or an antigen binding fragment thereof, comprising a heavy chain domain, wherein the heavy chain variable domain comprises a heavy chain complementarity determining region (HCDR)1, an HCDR2 and an HCDR3, and wherein the V H H monoclonal antibody specifically binds a Norovirus polypeptide, and wherein:
a) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 22, the CDR2 comprises amino acids 51-57 of SEQ ID NO:22, and the CDR3 comprises amino acids 96-110 of SEQ ID NO: 22;
b) the CDR1 comprises amino

Assignees

• The Us Secretary Depart Of Health And Human Services
• Instituto Nac De Tecnologia Agropecuaria
• Us Health
• Instituto Nacional De Tech Agropecuaria

Inventors

• Bok Karin
• Garaicoechea Lorena Laura
• Parreno Viviana
• Aguilar Andrea Pamela
• Bok Marina
• Green Lisbeth Kim
• Sosnovtsev Stanislav Vladimirovich

Classifications

• A61P31/14

  for RNA viruses · CPC title

• A61P31/12

  Antivirals · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• C07K2317/22

  from camelids, e.g. camel, llama or dromedary · CPC title

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title