Chloramphenicol acetyl transferase (CAT)-defective somatostatin fusion protein and uses thereof

• US 7,943,143 B2
• Filed: 02/10/2010
• Issued: 05/17/2011
• Est. Priority Date: 06/25/2008
• Status: Active Grant

• Current Assignee: Braasch Biotech LLC
• Original Assignee: Braasch Biotech LLC
• Inventors: Larrick, James, Mendelsohn, Andrew R., Haffer, Keith N.
• Primary Examiner(s): Graser; Jennifer E
• Application Number:
  US12/703,631
• Publication Number:
  US 20100136037A1
• Time in Patent Office: 461 Days
• Field of Search: None
• US Class Current: 424/198.1
• CPC Class Codes: A23K 50/60 for weanlings
• CPC Class Codes: A61K 2039/552 Veterinary vaccine
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 2039/55566 Emulsions, e.g. Freund's adjuvant, MF59
• CPC Class Codes: A61K 38/31 Somatostatins
• CPC Class Codes: A61K 39/0005 Vertebrate antigens from snakes A61K39/38
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/655 Somatostatins
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 9/1033 Chloramphenicol O-acetyltransferase (2.3.1.28)
• CPC Class Codes: C12Y 203/01028 Chloramphenicol O-acetyltransferase (2.3.1.28)

Abstract

Chimeric somatostatin-based polypeptides, polynucleotides used to encode the polypeptides, the methods for isolating and producing the polypeptides and the uses thereof are provided. In addition, low cost adjuvants for enhanced immunogenic response are provided. Vaccinations that include both chimeric somatostatin-based polypeptides and novel adjuvants are included, useful in facilitating farm animal productivity.

Claims

1. A method for increasing milk production in dairy cows comprising:

vaccinating a dairy cow with one or more doses of an immunogenic composition comprising;

(a) a chimeric polypeptide having the immunogenicity of somatostatin comprising an amino acid sequence of somatostatin-14 represented by SEQ ID NO;

1 linked to a substantially inactive and truncated chloramphenicol acetyl transferase polypeptide having a sequence selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

8, and SEQ ID NO;

9, wherein the somatostatin-14 is linked to the inactive chloramphenicol acetyl transferase by a spacer; and

(b) a pharmaceutically suitable adjuvant in an amount effective to elicit an immune response; and

allowing for at least ten days during which a dairy cow'"'"'s milk production will increase as compared to the same cow'"'"'s milk production in the absence of the vaccination.

2. The method of claim 1 wherein the dairy cow is vaccinated with only one dose of the immunogenic composition.

5. The method of claim 1, wherein the spacer comprises a sequence selected from the group consisting of SEQ ID NO:

11 and SEQ ID NO;

12.

6. The method of claim 1, wherein the chimeric polypeptide has an amino acid sequence with at least 99% sequence identity with the amino acid sequence of SEQ ID NO:

14.

7. The method of claim 1, wherein the chimeric polypeptide has an amino acid sequence of SEQ ID NO:

14.

8. The method of claim 1, wherein the pharmaceutically suitable adjuvant comprises an oil-in-water emulsion admixed with an emulsion premix wherein the oil-in-water emulsion comprises mineral oil, Tween 80, Span 85 and one or more polymers.

9. The method of claim 8, wherein the emulsion premix comprises a high molecular weight polymer, a surfactant, and an emulsifier in an oil-aqueous base.

10. The method of claim 1 wherein the dairy cow is vaccinated with only one dose of the immunogenic composition.

3. A method for increasing milk production in dairy cows comprising:

vaccinating a dairy cow with one or more doses of a composition comprising;

(a) a chimeric polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

8, and SEQ ID NO;

9 linked to an amino acid sequence of SEQ ID NO;

1, by a spacer; and

(b) an immunological adjuvant comprising;

an oil-in-water emulsion admixed with an emulsion premix wherein the oil-in-water emulsion comprises mineral oil, Tween 80, Span 85 and one or more polymers and wherein the emulsion premix comprises a high molecular weight polymer, a surfactant, and an emulsifier in an oil-aqueous base; and

allowing for at least ten days during which a dairy cow'"'"'s milk production will increase as compared to the same cow'"'"'s milk production in the absence of the vaccination.

4. A method for increasing lean meat production in a farm animal comprising:

vaccinating the farm animal with one or more doses of an immunogenic composition comprising;

(a) a chimeric polypeptide having the immunogenicity of somatostatin comprising an amino acid sequence of somatostatin-14 represented by SEQ ID NO;

1 linked to a substantially inactive and truncated chloramphenicol acetyl transferase polypeptide having a sequence selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

8, and SEQ ID NO;

9, wherein the somatostatin-14 is linked to the inactive chloramphenicol acetyl transferase by a spacer; and

(b) a pharmaceutically suitable adjuvant in an amount effective to elicit an immune response; and

allowing for several weeks during which the farm animal lean meat production will increase relative to lean meat production in a similar farm animal in the absence of the vaccination.

11. The method of claim 4, wherein the spacer comprises a sequence selected from the group consisting of SEQ ID NO:

11 and SEQ ID NO;

12.

12. The method of claim 4, wherein the chimeric polypeptide has an amino acid sequence with at least 99% sequence identity with the amino acid sequence of SEQ ID NO:

14.

13. The method of claim 4, wherein the chimeric polypeptide has an amino acid sequence of SEQ ID NO:

14.

14. The method of claim 4, wherein the pharmaceutically suitable adjuvant comprises an oil-in-water emulsion admixed with an emulsion premix wherein the oil-in-water emulsion comprises mineral oil, Tween 80, Span 85 and one or more polymers.

15. The method of claim 14, wherein the emulsion premix comprises a high molecular weight polymer, a surfactant, and an emulsifier in an oil-aqueous base.

16. The method of claim 4 wherein the farm animal is vaccinated with only one dose of the immunogenic composition.