# US 8,771,670 B2 · Identification of protective antigenic determinants of porcine reproductive and respiratory syndrome virus and uses thereof

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- Number: US 8,771,670 B2
- Document ID: US8771670B2
- Status: Granted
- Kind: B2
- Application number: 13/533,167
- Filed: 2012-06-26
- Published: 2013-06-20
- Granted: 2014-07-08
- Earliest claimed: 2005-11-29
- Assignees: Iowa State University Research Foundation, Inc.
- Applicants: Delbert Linn Harris; Matthew M. Erdman
- Inventors: Delbert Linn Harris; Matthew M. Erdman
- Web page: https://patentgrep.com/patent/US8771670B2

## CPC

- C07K 14/005: from viruses
- A61K 39/12: Viral antigens
- A61K 39/295: Polyvalent viral antigens (vaccinia virus or variola virus A61K39/285); Mixtures of viral and bacterial antigens
- A61P 31/12: Antivirals
- A61P 31/14: for RNA viruses
- A61P 37/04: Immunostimulants
- G01N 33/56983: {Viruses}
- A61K 2039/5252: {inactivated (killed)}
- A61K 2039/5254: {avirulent or attenuated}
- A61K 2039/552: {Veterinary vaccine}
- C12N 2770/10022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2770/10034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- 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)
- 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
- 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

The invention relates to a polypeptide of a protective antigenic determinant (PAD polypeptide) of porcine reproductive and respiratory syndrome virus (PRRSV) and nucleic acids encoding a PAD polypeptide. The PAD polypeptide and nucleic acids encoding a PAD polypeptide are useful in the development of antibodies directed to PAD, vaccines effective in providing protection against PRRSV infection, and diagnostic assays detecting the presence of PAD antibodies generated by a PAD-specific vaccine. The invention also discloses methods of generating antibodies to PAD, for vaccinating a pig to provide protection from PRRSV infections, a method of preparing the vaccine, a method of treating PRRSV infections in a pig, and a method of detecting antibodies to PAD of PRRSV.

## Claims

### Claim 1

A method for treating a porcine reproductive respiratory syndrome virus—(PRRSV) infection in a pig comprising:
administering to said pig a therapeutically effective amount of a vaccine comprising a first heterodimer of a matrix protein (M protein) of porcine reproductive and respiratory syndrome virus (PRRSV) and a glycoprotein 5 (GP5) of PRRSV (GP5-M heterodimer), wherein the GP5 of the first GP5-M heterodimer has glycosylation at position 44 of the GP5 in a North American (NA) PRRSV or glycosylation at position 46 of the GP5 in a European (EU) PRRSV; and
administering to said pig a second GP5-M heterodimer, wherein the GP5 of the second GP5-M heterodimer does not have glycosylation at position 44 of GP5 in a North American (NA) PRRSV or at position 46 of the GP5 in a European (EU) PRRSV, thereby treating a PRRSV infection in a pig, wherein the position number of 44 is determined from sequence alignment with the amino acid sequence of SEQ ID NO: 1, and the position number 46 is determined from the amino acid sequence of SEQ ID NO: 4.

### Claim 2

A multivalent vaccine comprising GP5-M protein heterodimers of PRRSV:
comprising a heterodimer of a matrix protein (M protein) of porcine reproductive and respiratory syndrome virus (PRRSV) and a glycoprotein 5 (GP5) of PRRSV, wherein said GP5 protein has N-glycosylation of asparagine amino acid located at position 44 of the GP5 protein in North American PRRSV strains or at position 46 of the GP5 protein in European PRRSV strains; and
comprising a heterodimer of a matrix protein (M protein) of porcine reproductive and respiratory syndrome virus (PRRSV) and a glycoprotein 5 (GP5) of PRRSV, wherein said GP5 protein does not have N-glycosylation of asparagine amino acid located at position 44 of the GP5 protein in North American PRRSV strains or at position 46 of the GP5 protein in European PRRSV strains, wherein the position number of 44 is determined from sequence alignment with the amino acid sequence of SEQ ID NO: 1, and the position number 46 is determined from the amino acid sequence of SEQ ID NO: 4.

### Claim 3

A GP5-M heterodimer subunit vaccine comprising:
comprising a heterodimer of a matrix protein (M protein of porcine reproductive and respiratory syndrome virus (PRRSV) and a glycoprotein 5 (GP5) of PRRSV, wherein said GP5 protein has N-glycosylation of asparagine amino acid located at position 44 of the GP5 protein in North American PRRSV strains or at position 46 of the GP5 protein in European PRRSV strains; and
comprising a heterodimer of a matrix protein (M protein) of porcine reproductive and respiratory syndrome virus (PRRSV) and a glycoprotein 5 (GP5) of PRRSV, wherein said GP5 protein does not have N-glycosylation of asparagine amino acid located at position 44 of the GP5 protein in North American PRRSV strains or at position 46 of the GP5 protein in European PRRSV strains, wherein the position number of 44 is determined from sequence alignment with the amino acid sequence of SEQ ID NO: 1, and the position number 46 is determined from the amino acid sequence of SEQ ID NO: 4.

### Claim 4 (depends on 3)

The subunit vaccine of claim 3 wherein the vaccine further comprises a heterodimer of said M protein of PRRSV and said GP5 of PRRSV, wherein the GP5 protein is linked to the M protein by a disulfide bond, wherein the disulfide bond results from bond between a cysteine amino acid in the M protein located at position 9 in North American PRRSV strains or at position 8 in EU PRRSV strains with a cysteine amino acid at position 48 of the GP5 protein in North American PRRSV strains or at position 50 in European PRRSV strains, thereby producing a GP5-M heterodimer, wherein the amino acid positions 9 and 48 in North American PRRSV strains are relatively to the amino acid reference sequence to the SEQ ID NO: 1 and the amino acid position of 8 and 50 in European PRRSV strains are relative to the reference amino acid sequence of SEQ ID NO: 4.

### Claim 5 (depends on 3)

The subunit vaccine of claim 3 wherein said GP5 further comprises at least one glycosylated amino acid from amino acids 1-43 of the GP5 ectodomain in North American PRRSV strains or from amino acids 1-45 of the GP5 ectodomain in European PRRSV strains, wherein the amino acids 1-43 of North American PRRSV strains are relatively to the amino acid reference sequence to the SEQ ID NO: 1 and amino acids position 1-45 in European PRRSV strains are relative to the reference amino acid sequence of SEQ ID NO: 4.