# US 11,307,202 B1 · Antibody binding detection method for detecting MERS-CoV

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,307,202 B1
- Document ID: US11307202B1
- Status: Granted
- Kind: B1
- Application number: 17/406,182
- Filed: 2021-08-19
- Granted: 2022-04-19
- Earliest claimed: 2021-01-29
- Assignees: King Abdulaziz University
- Applicants: King Abdulaziz University
- Inventors: Ayman Talaat Abbas Abdelhadi; Esam Ibraheem Ahmed Azhar; Sherif Aly Abdelkhalek Elkafrawy; Sayed Sartaj Sohrab
- Web page: https://patentgrep.com/patent/US11307202B1

## CPC

- A61K 39/12: Viral antigens
- A61K 39/215: Coronaviridae, e.g. avian infectious bronchitis virus
- A61P 31/14: for RNA viruses
- C07K 16/102: Coronaviridae (F)
- G01N 33/54386: {Analytical elements}
- G01N 33/56983: {Viruses}
- A61K 2039/55566: {Emulsions, e.g. Freund's adjuvant, MF59}
- C07K 2317/11: isolated from eggs
- C07K 2317/23: from birds
- C07K 2317/30: characterized by aspects of specificity or valency
- C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
- C12N 2770/20011: Coronaviridae
- C12N 2770/20034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- G01N 2333/165: Coronaviridae, e.g. avian infectious bronchitis virus
- G01N 2469/10: Detection of antigens from microorganism in sample from host
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A method for detecting MERS-CoV at high sensitivity and specificity using IgY antibodies that bind to MERS-CoV N protein, its fragments and domains. Isolated or purified IgY monospecific antibodies to MERS-CoV N protein.

## Claims

### Claim 1

An antibody binding method for detecting MERS-CoV in Camelidae, comprising
obtaining a nasopharyngeal swab sample from a subject,
contacting the nasopharyngeal swab sample with a monospecific IgY antibody that recognizes MERS-CoV nucleocapsid protein (“N protein”), wherein the contacting includes contacting the sample with a calcium alginate swab to which the monospecific IgY antibody is bound, then removing a remaining unbound portion of the nasopharyngeal swab sample, and
detecting complex formation or binding of the IgY antibody to the MERS-CoV,
wherein complex formation or binding is detected with an ELISA sandwich assay, wherein the IgY antibody that recognizes MERS-CoV N protein is a capturing antibody bound to a substrate, wherein the capturing antibody bound to the substrate is contacted with the sample, and wherein complex formation is measured by the binding of a tagged secondary antibody that recognizes MERS-CoV,
wherein the substrate is a microtiter plate well or a chromatography bead,
wherein the tagged secondary antibody is bioconjugated to an enzyme.

### Claim 2 (depends on 1)

The method of claim 1, wherein said sample contains no more than 20 ng/ml of MERS-CoV N protein.

### Claim 3 (depends on 1)

The method of claim 1, wherein the IgY antibody is a polyclonal antibody produced by immunizing chickens with isolated or recombinant MERS-CoV N protein and recovering IgY antibody from eggs laid by the immunized chickens.

### Claim 4 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a polyclonal monospecific IgY antibody for MERS-CoV N protein.

### Claim 5 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a monospecific antibody that binds to a peptide epitope of MERS-CoV N protein that ranges in length from 6 to 20 amino acids of the amino acid sequence described by SEQ ID NO: 1.

### Claim 6 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a monospecific antibody that binds to an N-terminal domain segment of MERS CoV N protein comprising residues 1-36 of SEQ ID NO: 1.

### Claim 7 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a monospecific antibody that binds to an N-terminal domain segment of MERS CoV N protein comprising residues 37-164 of SEQ ID NO: 1.

### Claim 8 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a monospecific antibody that binds to a central segment of MERS CoV N protein comprising residues 165-238 of SEQ ID NO: 1; or that binds to a disordered region or LKR containing a stretch of serine and arginine residues and comprising residues 156 to 206 of SEQ ID NO: 1 or LPKNFHIEGTGGNSQSSSRASSVSRNSSRSSSQGSRSGNSTRGTSPGPSGI (SEQ ID NO: 3).

### Claim 9 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a monospecific antibody that binds to a C-terminal domain segment of MERS CoV N protein comprising residues 239-362 of SEQ ID NO: 1.

### Claim 10 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a monospecific antibody that binds to a C-terminal domain segment of MERS CoV N protein comprising residues 363-413 of SEQ ID NO: 1.

### Claim 11 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a monospecific antibody that binds to one or more residues in a peptide segment of SEQ ID NO: 1 selected from the group comprising g residues 4-12, 42-47, 50-58, 60-66, 92-102, 108-126, 144-150, 172-180, 206-225, 230-240, 261-268, 296-304, 324-337, 347-353 and 399-405 described by SEQ ID NO: 1.

### Claim 12 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a monospecific antibody that binds to one or more residues in a peptide segment of SEQ ID NO: 1 selected from the group comprising:
K346, W347, L348 and E349;
Y327, F328, L329, and R330;
L350, L351, E352, O353, N354, 1355, D356, A357, Y358, K359, T360, F361, P362, K363, K364, and E365;
D320, D321, H322, G323, N324, P325, and V326;
H252, K253, R254 T255, S256, T257, K258, S259, F260, N261 M262, V263, Q264, A265, F266, G267, L268, R269, G270, P271, G272, D273, L274, Q275, G276, N277, F278, G279, D280, L281, Q282, L283, N284, K285, L286, G287, T288, E289, D290, P291, and R292;
R399, P400, S401, V402, Q403, P404, G405, P406, M407, 1408, D409, V410, N411, T412, and D413;
W293, P294, 0295, 1296, A297, and E298;
F312, K313, L314, T315, H316, Q317, N318, and N319; and
L299, A300, P301, T302, A303, S304, A305, F306, M307, G308, M309, S310, and Q311.

### Claim 13 (depends on 1)

The method of claim 1, wherein the IgY antibody comprises a mixture of two or more monospecific IgY antibodies to MERS CoV N protein epitopes or domains.

### Claim 14 (depends on 1)

The method of claim 1, wherein the tagged secondary antibody is a peroxidase-conjugated anti-rabbit antibody.