# US 9,897,605 B2 · Porcine torque teno virus vaccines and diagnosis

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- Number: US 9,897,605 B2
- Document ID: US9897605B2
- Status: Granted
- Kind: B2
- Application number: 14/946,384
- Filed: 2015-11-19
- Published: 2016-07-28
- Granted: 2018-02-20
- Earliest claimed: 2009-08-21
- Assignees: Virginia Tech Intellectual Properties, Inc.
- Applicants: Virginia Tech Intellectual Properties, Inc.
- Inventors: Xiang-Jin Meng; Yaowei Huang
- Web page: https://patentgrep.com/patent/US9897605B2

## CPC

- G01N 33/56983: {Viruses}
- A61K 39/12: Viral antigens
- A61P 31/20: for DNA viruses
- A61P 37/04: Immunostimulants
- C07K 14/005: from viruses
- C07K 16/081: DNA viruses
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- C12Q 1/701: {Specific hybridization probes}
- A61K 2039/552: {Veterinary vaccine}
- C12N 2750/00021: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- C12N 2750/00022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2750/00034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2750/14034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- G01N 2333/01: DNA viruses
- G01N 2333/085: Picornaviridae, e.g. coxsackie virus, echovirus, enterovirus
- G01N 2469/20: Detection of antibodies in sample from host which are directed against antigens from microorganisms
- 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)
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present invention provides four purified preparation containing a polynucleic acid molecule encoding porcine Torque teno virus (PTTV) genotypes or subtypes PTTV1a-VA, PTTV1b-VA, PTTV2b-VA, and PTTV2c-VA. The present invention also provides infectious DNA clones, biologically functional plasmid or viral vector containing the infectious nucleic acid genome molecule of the same. The present invention further provides live, attenuated, vector-expressed and purified recombinant capsid subunit or killed viral vaccines for protection against PTTV infection. The present invention additionally provides subunit vaccines comprising PTTV specific gene products, especially ORF1 capsid gene product for protection against PTTV infection. Further, the present invention provides methods for diagnosing PTTV infection via polymerase chain reaction (PCR) using specific primer for PTTV1, PTTV2, and individual PTTV1 genotypes. Finally, the present invention provides methods for diagnosing PTTV infection via immunological methods, e.g., enzyme-linked immunoabsorbent assay (ELISA) and Western blot using PTTV specific antigens for detecting serum PTTV specific antibodies.

## Claims

### Claim 1

A vaccine comprising at least one adjuvant in an immunologically effective amount and a Porcine Torque Teno Virus open reading frame 1 (ORF1) protein according to SEQ ID NO: 16, or an immunogenic fragment of the ORF1 protein according to SEQ ID NO: 16 that comprises subtype specific hypervariable regions (HVR) represented by amino acids 363 to 375 and 388 to 423 of SEQ ID NO: 16.

### Claim 2 (depends on 1)

The vaccine according to claim 1, wherein ORF1 protein or immunogenic fragment of the ORF1 protein is a purified plasmid or baculovirus vector expressed recombinant protein.

### Claim 3 (depends on 1)

The vaccine according to claim 1, further comprising at least one physiologically acceptable carrier.

### Claim 4 (depends on 1)

The vaccine according to claim 1, wherein the adjuvant is selected from one or more of aluminum hydroxide (alum), aluminum potassium sulfate, immunostimulating complexes (ISCOMS), non-ionic block polymers or copolymers, cytokines, saponins, monophosphoryl lipid A (MLA), muramyl dipeptides (MDP), enterotoxins isolated from Escherichia coli, cholera toxins, diphtheria toxin, tetanus toxin, pertussis toxin, and subunits of cholera, diphtheria, tetanus, and pertussis toxins.

### Claim 5

A vaccine for protecting against a Porcine Torque Teno Virus (PTTV) infection, wherein the vaccine comprises at least one adjuvant and a recombinant protein comprising amino acids 310 to 625 of SEQ ID NO: 16.

### Claim 6 (depends on 1)

The vaccine according to claim 1, which is adapted to be administered parenterally, intranasally, intradermally, or transdermally to a pig.

### Claim 7 (depends on 1)

The vaccine according to claim 1, which is adapted to be administered intralymphoidly or intramuscularly to a pig.

### Claim 8

An immunogenic composition comprising at least one adjuvant and a recombinant subunit capsid protein or immunogenic fragment thereof expressed from a bacterial or baculovirus expression system wherein the capsid protein or immunogenic fragment thereof comprises a Porcine Torque Teno Virus polypeptide represented by amino acids 363 to 375 of SEQ ID NO: 16 and a Porcine Torque Teno Virus polypeptide represented by amino acids 388 to 423 of SEQ ID NO: 16.

### Claim 9 (depends on 8)

The immunogenic composition according to claim 8, which is adapted to be administered parenterally, intranasally, intradermally, transdermally, intralymphoidly or intramuscularly to a pig.

### Claim 10

A plasmid or baculovirus vector encoding a recombinant capsid protein or immunogenic fragment thereof wherein the recombinant capsid protein or immunogenic fragment comprises a Porcine Torque Teno Virus polypeptide represented by amino acids 363 to 375 of SEQ ID NO: 16 and a Porcine Torque Teno Virus polypeptide represented by amino acids 388 to 423 of SEQ ID NO: 16.

### Claim 11 (depends on 10)

The plasmid or baculovirus vector of claim 10, wherein the recombinant capsid protein or immunogenic fragment thereof comprises amino acids 310 to 625 of SEQ ID NO: 16.