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

• US 8,367,073 B2
• Filed: 03/23/2011
• Issued: 02/05/2013
• Est. Priority Date: 06/25/2008
• Status: Active Grant

• Current Assignee: Braasch Biotech LLC
• Original Assignee: Braasch Biotech LLC
• Inventors: Mendelsohn, Andrew R., Haffer, Keith N., Larrick, James
• Primary Examiner(s): GRASER, JENNIFER E
• Application Number:
  US13/069,592
• Publication Number:
  US 20120076806A1
• Time in Patent Office: 685 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 composition comprising:

a chimeric polypeptide comprising a somatostatin-14 polypeptide represented by SEQ ID NO;

1 and a 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 composition is substantially endotoxin free.

2. The composition of claim 1, wherein the composition is at least 80% endotoxin free.

3. The composition of claim 1, wherein the chimeric polypeptide has an amino acid sequence with at least 90% sequence identity to SEQ ID NO:

14.

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

14.

5. The composition of claim 1, formulated for use as a vaccine.

6. The composition of claim 1, wherein the chloramphenicol acetyl transferase polypeptide lacks enzymatic activity.

7. The composition of claim 1, wherein the somatostatin-14 polypeptide and the inactive chloramphenicol acetyl transferase polypeptide are linked by a spacer.

8. The composition of claim 7, wherein the spacer comprises a sequence selected from the group consisting of SEQ ID NO:

11 or SEQ ID NO;

12.

9. The composition of claim 1, further comprising an adjuvant.

10. The composition of claim 9, further comprising one or more antigens selected from the group consisting of native polypeptides, recombinant polypeptides, synthetic polypeptides, polysaccharides, and glycoproteins.

11. A composition comprising:

a chimeric polypeptide comprising a somatostatin-14 polypeptide represented by SEQ ID NO;

1 and a chloramphenicol acetyl transferase polypeptide,

wherein the chloramphenicol acetyl transferase polypeptide is modified at the 192 residue, at the 193 residue, or at both 192 and 193 residues, relative to the wild type chloramphenicol acetyl transferase polypeptide represented by residues 1-210 of SEQ ID NO;

15, and

wherein the modification comprises replacement of one or both histidines independently with glycine or alanine.

12. The composition of claim 11, wherein the modification comprises replacement of histidine at the 193 residue to a glycine.

13. The composition of claim 11, wherein the modification comprises replacement of histidine at the 193 residue to an alanine.

14. The composition of claim 11, wherein the modification comprises replacement of histidine at the 192 residue to a glycine and histidine at the 193 residue to a glycine.

15. The composition of claim 11, wherein the chloramphenicol acetyl transferase polypeptide is truncated at the C-terminus by 10 amino acids relative to the wild type polypeptide.

16. The composition of claim 11, wherein the composition is substantially endotoxin free.

17. The composition of claim 11, wherein the composition is at least 80% endotoxin free.