Chimeric somatostatin containing protein and encoding DNA, plasmids of expression, method for preparing chimeric protein, strain-producers, immunogenic composition, method for increasing the productivity of farm animals

• US 6,316,004 B1
• Filed: 06/22/1994
• Issued: 11/13/2001
• Est. Priority Date: 06/22/1993
• Status: Expired due to Fees

• Current Assignee: T Tikhonenko
• Original Assignee: T Tikhonenko
• Inventors: Lunin, Vladimir Glebovich, Karpov, Vladimir Abramovich, Khodun, Marat-Vladimir Leonidovich, Sergienko, Olga Vasileivna, Tikhonenko, Tomas Iosifovich, Bader, Leila Bakievna
• Primary Examiner(s): Achutamurthy, Ponnathapura
• Assistant Examiner(s): Bui, Phuong T.
• Application Number:
  US08/264,042
• Time in Patent Office: 2,701 Days
• Field of Search: 530/311, 435/69.4, 424/198.1, 424/185.1
• US Class Current: 424/198.1
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: C07K 14/655 Somatostatins
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 9/003 Dihydrofolate reductase [DHFR] (1.5.1.3)
• CPC Class Codes: C12N 9/1033 Chloramphenicol O-acetyltransferase (2.3.1.28)

Abstract

Chimeric polypeptides having the immunogenicity of somatostatin include an amino acid sequence of somatostatin-14 and a protein carrier. The sequence of somatostatin-14 is joined to the 3′-end of the protein carrier by a spacer (Sp)n, wherein Sp consists of an alkaline amino acid and an amino acid that provides a rigid expended chain-like β-structure, and n designates the number of blocks in the spacer. In a preferred embodiment, n is from 1 to 8 and the protein-carrier is chloramphenicol acetyl transferase (CAT).

Claims

1. A chimeric polypeptide having the immunogenicity of somatostatin, including an amino acid sequence of somatostatin-14 and a protein carrier, wherein the sequence of somatostatin-14 is joined to the 3′

-end of the protein carrier through a spacer (Sp)_n, where Sp consists of an alkaline amino acid and an amino acid providing a rigid β

-structure, and n designates the number of blocks in the spacer, wherein the polypeptide is encoded by Seq. ID No. 1 or Seq. ID No. 2, wherein Sp is Arg—

Pro or Lys—

Pro, and wherein n=4.

2. The polypeptide of claim 1, wherein the protein-carrier is chloramphenicol acetyl transferase.

3. The polypeptide of claim 2, wherein the chloramphenicol acetyl transferase has a deletion 10 amino acids long at the C-end.

4. The polypeptide of claim 1, wherein the protein carrier is dihydrofolatereductase.

5. The polypeptide of claim 2, where said polypeptide is water insoluble.

6. An immunogenic composition comprising the chimeric polypeptide of claim 1 together with a pharmaceutically suitable adjuvant in an amount effective to elicit an immune response.

7. The composition of claim 6, wherein the adjuvant is Freund'"'"'s incomplete adjuvant.

8. The composition of claim 7, wherein the weight ratio of purified chimeric somatostatin-comprising protein to incomplete Freund'"'"'s adjuvant is from 1:

0.5 to 1;

10.

9. The composition of claim 8, wherein the polypeptide and adjuvant are in a ration of 1:

1.

10. The polypeptide of claim 3 encoded by Seq. ID No. 1 wherein Sp is Arg—

Pro and n=4.

11. The polypeptide of claim 3 encoded by Seq. ID No. 2 wherein Sp is Lys—

Pro and n=4.

12. The chimeric polypeptide of claim 1 encoded by a recombinant plasmid and being expressed under the control of constitutive or inducible promoters in bacterial cells.

13. The chimeric polypeptide of claim 12 wherein said bacterial cells comprise protease deficient E. coli strains.