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

Patent metadata

Field	Value
Publication number	US-11325965-B2
Application number	US-202016854703-A
Country	US
Kind code	B2
Filing date	Apr 21, 2020
Priority date	May 9, 2013
Publication date	May 10, 2022
Grant date	May 10, 2022

Abstract

Isolated VHH 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. A method for detecting Norovirus in a sample, comprising:
contacting the sample with an isolated V H H monoclonal antibody or an antigen binding fragment thereof; and
detecting the V H H monoclonal antibody bound to the sample,
wherein the presence of the V H H monoclonal antibody bound to the sample indicates the presence of the Norovirus,
wherein the V H H monoclonal antibody specifically binds a Norovirus polypeptide and comprises a heavy chain variable domain, wherein the heavy chain variable domain comprises a heavy chain complementarity determining region (CDR)1, a CDR2 and a CDR3, 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 CDR3 comprises amino acids 96-109 of SEQ ID NO: 4;
e) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 5, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 5, and the CDR3 comprises 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 CDR3 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 of SEQ ID NO: 11, 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 comprises amino acids 26-33 of SEQ ID NO: 18, the CDR2 comprises amino acids 51-58 of SEQ ID NO: 18, and the CDR3 comprises 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 comprises 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;
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;
v) 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;
w) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 23, the CDR2 comprises amino acids 50-56 of SEQ ID NO:23, and the CDR3 comprises amino acids 95-104 of SEQ ID NO: 23;
x) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 24, the CDR2 comprises amino acids 51-57 of SEQ ID NO:24, and the CDR3 comprises amino acids 96-107 of SEQ ID NO: 24;
y) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 25, the CDR2 comprises amino acids 51-57 of SEQ ID NO: 25, and the CDR3 comprises amino acids 100-110 of SEQ ID NO: 25;
z) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 26, the CDR2 comprises amino acids 50-57 of SEQ ID NO:26, and the CDR3 comprises amino acids 96-117 of SEQ ID NO: 26;
aa) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 27, the CDR2 comprises amino acids 51-58 of SEQ ID NO:27, and the CDR3 comprises amino acids 97-108 of SEQ ID NO: 27;
bb) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 28, the CDR2 comprises amino acids 51-57 of SEQ ID NO:28, and the CDR3 comprises amino acids 96-112 of SEQ ID NO: 28;
cc) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 29, the CDR2 comprises amino acids 51-58 of SEQ ID NO:29, and the CDR3 comprises amino acids 97-115 of SEQ ID NO: 29; or
dd) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 30, the CDR2 comprises amino acids 51-58 of SEQ ID NO:30, and the CDR3 comprises amino acids 97-121 of SEQ ID NO: 30,
thereby detecting the Norovirus in the sample.

2. The method of claim 1 , wherein the V H H monoclonal antibody or an antigen binding fragment thereof is directly labeled.

3. The method of claim 1 , further comprising:
contacting the sample with a second antibody that specifically binds the V H H monoclonal antibody or an antigen binding fragment thereof; 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 the Norovirus in the sample.

4. The method of claim 1 , wherein the Norovirus is Genogroup I Norovirus.

5. The method of claim 1 , wherein the Norovirus is a Genogroup II Norovirus.

6. The method of claim 1 , 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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 CDR3 comprises amino acids 96-109 of SEQ ID NO: 4, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 4;
e) the CDR1 comprises amino acids 26-33 of SEQ ID NO: 5, the CDR2 co

Assignees

• Us Health
• Instituto Nac De Tecnologia Agropecuaria
• 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

• C07K16/10 Primary

  RNA viruses · CPC title

• G01N33/56983 Primary

  Viruses · CPC title

• C07K2317/76

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

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C12N2770/16034

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title