IP Library › Granted Patent US 11,325,965

Granted Patent B2

US 11,325,965 · App. 16/854,703 · Granted May 10, 2022

Single-domain V

Inventors: Karin Bok (Bethesda, MD); Lorena Laura Garaicoechea (Buenos Aires, AR); Viviana Parreno (Buenos Aires, AR); Andrea Pamela Aguilar (Buenos Aires, AR); Marina Bok (Buenos Aires, AR); Lisbeth Kim Green (Olney, MD); Stanislav Vladimirovich Sosnovtsev (North Potomac, MD)

Assignees: The United States of America, as represented by the Secretary, Department of Health and Human Services; Instituto Nacional De Technologia Agropecuaria

C07K16/10 G01N33/56983 A61K2039/505 C07K2317/22 C07K2317/24 C07K2317/33 C07K2317/34 C07K2317/565 C07K2317/569 C07K2317/76 C07K2317/92 C12N2770/16011 C12N2770/16034 G01N2333/08

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Quick Facts

Patent No.: US 11,325,965
App. No.: 16/854,703
Granted: May 10, 2022
Kind: B2

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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.

Claims (157)

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 comprises amino acids 51-58 of SEQ ID NO: 5, and the CDR3 comprises amino acids 97-110 of SEQ ID NO: 5, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 21.

7. The method of claim 1 , 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 22;

b) 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 23;

c) 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 24;

d) 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 25;

e) 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 26;

f) 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 27;

g) 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 28;

h) 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 29; or

i) 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, and wherein the heavy chain variable domain of the V H H monoclonal antibody is at least 90% identical to SEQ ID NO: 30.

8. The method of claim 6 , wherein the heavy chain variable domain comprises the amino acid sequence set for the as one of SEQ ID NOs: 1-21.

9. The method of claim 7 , wherein the heavy chain variable domain comprises the amino acid sequence set forth as one of SEQ ID NOs: 22-30.

10. The method of claim 1 , wherein the sample is a tissue, blood, serum, plasma, spinal fluid, sputum, nasopharyngeal secretion, stool or urine sample.

11. The method of claim 1 , wherein the sample is an environmental sample.

12. The method of claim 1 , wherein the sample comprise a subunit vaccine.

13. The method of claim 2 , wherein the label is an enzyme, prosthetic group, fluorescent material, luminescent material, magnetic agent or radioactive material.

14. A kit for detecting a Norovirus, comprising:

i) a container comprising an isolated V H H monoclonal antibody or an antigen binding fragment thereof, 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: