DNA encoding and methods of production of insulin-like growth factor binding protein BP53

• US 5,258,287 A
• Filed: 03/22/1988
• Issued: 11/02/1993
• Est. Priority Date: 03/22/1988
• Status: Expired due to Term

• Current Assignee: Royal Prince Alfred Hospital (Australian Unity Limited), Central Sydney Area Health Service, Genetec Incorporated (Noritsu Koki Co., Ltd.)
• Original Assignee: Central Sydney Area Health Service, Genentech, Inc. (Roche Holding AG)
• Inventors: Baxter, Robert C., Wood, William I.
• Primary Examiner(s): Hill, Jr., Robert J.
• Assistant Examiner(s): Allen, Marianne Porta
• Application Number:
  US07/171,623
• Time in Patent Office: 2,051 Days
• Field of Search: 435/69.1, 435/6, 435/172.3, 435/69.8, 435/320.1, 435/254, 435/255, 435/256, 435/240.2, 435/240.1, 536/27
• US Class Current: 435/69.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4743 Insulin-like growth factor binding protein
• CPC Class Codes: C07K 14/755 Factors VIII , e.g. factor VIII C (AHF), factor VIII Ag (VWF)

Abstract

DNA isolates coding for insulin-like growth factor binding protein may be used to produce the protein via recombinant expression systems. Insulin-like growth factor binding protein, which generally has a molecular weight of about 53 kD on non-reducing SDS-PAGE, is useful as a binder to insulin-like growth factor and as a metabolic regulator.

Claims

1. An isolated DNA molecule comprising a sequence that hybridizes, under stringent conditions of 50% formamide with 0.75M NaCl and 0.075M sodium citrate, at 42°

C., to the portion of the DNA sequence of FIG. 3 coding for mature BP53 or the preprotein for BP53 and which encodes a BP53 protein that binds to IGF-I or IGF-II, excluding BP28, PP12, and HEP-G2.

2. The DNA molecule of claim 1 further comprising a promoter operably linked to said protein-encoding sequence.

3. The DNA molecule of claim 1 further comprising an origin of replication operative in a unicellular organism.

4. The molecule of claim 1 further comprising a signal sequence-encoding region upstream from and operably linked to the protein-encoding sequence.

5. The molecule of claim 4 wherein the signal sequence is recognized by mammalian host cells.

6. The DNA molecule of claim 1 wherein the BP53 protein is human or porcine BP53.

7. An expression vector for producing an insulin-like growth factor binding protein comprising the DNA molecule of claim 1 operably linked to control sequences for expression of the DNA sequence.

8. A host cell transformed with the expression vector of claim 7.

9. The host cell of claim 8 wherein the cell is prokaryotic.

10. The host cell of claim 8 that is eukaryotic.

11. The host cell of claim 10 that is mammalian.

12. The host cell of claim 11 that is a human embryonic kidney cell or COS cell.

13. The host cells of claim 8 that are from a multicellular organism.

14. The host cells of claim 8 that are from a vertebrate.

15. The host cells of claim 8 that are microbial cells.

16. The host cells of claim 8 that are bacterial, yeast fungal, insect, plant, or mammalian cells.

17. A method of using a DNA molecule encoding an insulin-like growth factor binding protein to produce said protein, which method comprises culturing the cells of claim 8 to express the DNA sequence encoding the protein.

18. The method of claim 17 further comprising the step of recovering the protein from the host cell culture.

19. A method of using a DNA molecule encoding an insulin-like growth factor binding protein to produce said protein, which method comprises culturing the cells of claim 10 to express the DNA sequence encoding the protein.

20. An isolated DNA molecule comprising the portion of the DNA sequence of FIG. 3 encoding mature BP53 or the preprotein of BP53.