# US 2014/0193829 A1 · Monoclonal antibodies against scrapie prion protein

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- Number: US 2014/0193829 A1
- Document ID: US20140193829A1
- Status: Published application
- Kind: A1
- Application number: 14/140,326
- Filed: 2013-12-24
- Published: 2014-07-10
- Earliest claimed: 2012-12-25
- Assignees: National Yang-Ming University
- Applicants: National Yang-Ming University
- Inventors: Yi-Ming Arthur CHEN; Szu-Wei HUANG; Ying-YU LIN
- Web page: https://patentgrep.com/patent/US20140193829A1

## CPC

- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- C07K 16/2872: {against prion molecules, e.g. CD230}
- G01N 33/6896: {Neurological disorders, e.g. Alzheimer's disease}
- C07K 2317/33: Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
- C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
- G01N 2800/2828: Prion diseases
- 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)
- 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)

## Abstract

The present invention relates to monoclonal antibodies against PrPSc protein. The anti-PrPSc mAbs produced of this invention can specifically detect PrPSc protein from normal mouse brain homogenates without pre-treatment with protease K, which may be used in developing a diagnostic method and kit for prion disease with high specificity and sensitivity.

## Claims

### Claim 1

A monoclonal antibody specific to scrapie prion protein (PrPSc), which binds to a peptide comprising the amino acid sequence of SEQ ID NO.1, composed of the amino acid residues 141-182 of prion protein.

### Claim 2 (depends on 1)

The monoclonal antibody of claim 1, wherein the monoclonal antibody specifically recognizes the β-sheet structure of PrPSc.

### Claim 3 (depends on 1)

The monoclonal antibody of claim 1, wherein the monoclonal antibody specifically recognizes the prion protein which is conformed to a β oligomer.

### Claim 4 (depends on 1)

The monoclonal antibody of claim 1, wherein the monoclonal antibody binds to a peptide comprising the amino acid sequence of SEQ ID NO.2, composed of the amino acid residues 141-152 of prion protein.

### Claim 5 (depends on 1)

The monoclonal antibody of claim 1, wherein the monoclonal antibody binds to a peptide comprising the amino acid sequence of SEQ ID NO.3, composed of the amino acid residues 171-182 of prion protein.

### Claim 6

A producing method of an antibody for specifically detecting scrapie prion protein (PrPSc), comprising:
preparing a recombinant prion protein with β-sheet structure (βPrP);
immunizing a mouse with the recombinant PrPSc to obtain an antibody against the β PrP; and
isolating and purifying the antibody.

### Claim 7 (depends on 6)

The method of claim 6, wherein the antibody includes a monoclonal antibody and a polyclonal antibody.

### Claim 8 (depends on 6)

The method of claim 6, wherein the β-sheet structure of the recombinant prion protein is refolded from the α-helical structures of PrPC, and further polymerized into a β oligomer.

### Claim 9 (depends on 6)

The method of claim 6, wherein the recombinant prion protein comprises 4 amino acid mutations at Ser-132, Asn-181, Cys179 and Cys-214.

### Claim 10 (depends on 1)

A diagnostic kit for specifically detecting scrapie prion protein (PrPSc), which is characterized by comprising the monoclonal antibody of claim 1.

### Claim 11 (depends on 1)

A method for the detection of prion diseases in animals or humans, comprising steps of:
(a) preparing a sample from brain tissue of the tested animal without protease treatment; and
(b) detecting the presence of infectious form of PrP (PrPSc) in the sample by a immunoassay using the monoclonal antibody of claim 1.

### Claim 12 (depends on 11)

The method of claim 11, wherein the brain tissue sample is prepared as a brain tissue homogenate.

### Claim 13 (depends on 12)

The method of claim 12, wherein the immunoassay is Western blot assay.

### Claim 14 (depends on 11)

The method of claim 11, wherein the brain tissue sample is prepared as a brain tissue slice.

### Claim 15 (depends on 14)

The method of claim 14, wherein the immunoassay is immunohistochemistry assay.

### Claim 16 (depends on 11)

The method of claim 11, wherein the infectious form of PrP comprises a β-sheet structure.

### Claim 17 (depends on 16)

The method of claim 16, wherein the infectious form of PrP is conformed to a β oligomer.