Claims
- 1. A purified and isolated polypeptide having part or all of the primary structural conformation and one or more of the biological properties of naturally-occurring pluripotent granulocyte colony-stimulating factor and characterized by being the product of procaryotic or eucaryotic expression of an exogenous DNA sequence.
- 2. A polypeptide according to claim 1 further characterized by being free of association with any mammalian protein.
- 3. A polypeptide according to claim 1 wherein the exogenous DNA sequence is a cDNA sequence.
- 4. A polypeptide according to claim 1 wherein the exogenous DNA sequence is a manufactured DNA sequence.
- 5. A polypeptide according to claim 1 wherein the exogenous DNA sequence is a genomic DNA sequence.
- 6. A polypeptide according to claim 1 wherein the exogenous DNA sequence is carried on an autonomously replicating DNA plasmid or viral vector.
- 7. A polypeptide according to claim 1 possessing part or all of the primary structural conformation of human pluripotent granulocyte colony-stimulating factor as set forth in Table VII or any naturally occurring allelic variant thereof.
- 8. A polypeptide according to claim 1 which has the immunological properties of naturally-occurring pluripotent granulocyte colony-stimulating factor.
- 9. A polypeptide according to claim 1 which has the in vitro biological activity of naturally-occurring pluripotent granulocyte colony-stimulating factor.
- 10. A polypeptide according to claim 1 further characterized by being covalently associated with a detectable label substance.
- 11. A polypeptide according to claim 10 wherein said detectable label is a radiolabel.
- 12. A DNA sequence for use in securing expression in a procaryotic or eucaryotic host cell of a polypeptide product having at least a part of the primary structural conformation and one or more of the biological properties of naturally-occurring pluripotent granulocte colony-stimulating factor, said DNA sequence selected from among:
(a) the DNA sequence set out in Tables VII and VII or their complimentary strands; (b) DNA sequences which hybridize to the DNA sequences defined in (a) or fragments thereof; and (c) DNA sequences which, but for the degeneracy of the genetic code, would hybridize to the DNA sequences defined in (a) and (b).
- 13. A procaryotic or eucaryotic host cell transformed or transfected with a DNA sequence according to claim 12 in a manner allowing the host cell to express said polypeptide product.
- 14. A polypeptide product of the expression of a DNA sequence of claim 12 in a procaryotic or eucaryotic host.
- 15. A purified and isolated DNA sequence coding for procaryotic or eucaryotic host expression of a polypeptide having part or all of the primary structural conformation and one or more of the biological properties of pluripotent granulocyte colony-stimulating factor.
- 16. A cDNA sequence according to claim 15.
- 17. A genomic DNA sequence according to claim 16.
- 18. A manufactured DNA sequence according to claim 15.
- 19. A manufactured DNA sequence according to claim 18 and including one or more codons preferred for expression in E. coli cells.
- 20. A manufactured DNA sequence according to claim 18, and coding for expression of human species pluripotent granulocyte colony-stimulating factor.
- 21. A manufactured DNA sequence according to claim 20 and including one or more codons preferred for expression in yeast cells.
- 22. A human species pluripotent granulocyte colony-stimulating factor-coding DNA sequence according to claims 16, 17 or 18.
- 23. A DNA sequence according to claim 15 covalently associated with a detectable label substance.
- 24. A DNA sequence according to claim 23 wherein the detectable label is a radiolabel.
- 25. A single-stranded DNA sequence according to claim 23.
- 26. A DNA sequence coding for a polypeptide fragment or polypeptide analog of naturally-occurring pluripotent granulocyte colony-stimulating factor.
- 27. A DNA sequence coding for [Ala1] hpG-CSF.
- 28. A biologically functional plasmid or viral DNA vector including a DNA sequence according to one of claims 12, 15 or 26.
- 29. A procaryotic or eucaryotic host cell stably transformed or transfected with a DNA vector according to claim 28.
- 30. A polypeptide product of the expression in a procaryotic or eucaryotic host cell of a DNA sequence according to claims 15 or 26.
- 31. A synthetic polypeptide having part or all of the amino acid sequence as set forth in Table VII and having one or more of the in vitro biological activities of naturally-occurring pluripotent granulocyte colony-stimulating factor.
- 32. A synthetic polypeptide having part or all of the secondary conformation of part or all of the amino acid sequence set forth in Table VII and having a biological property of naturally-occurring human pluripotent granulocyte colony-stimulating factor.
- 33. A process for the production of a polypeptide having part or all of the primary structural conformation and one or more of the biological properties of naturally occurring pluripotent granulocyte colony-stimulating factor, said process comprising:
growing, under suitable nutrient conditions, procaryotic or eucaryotic host cells transformed or transfected with a DNA vector according to claim 28, and isolating desired polypeptide products of the expression of DNA sequences in said vector.
- 34. Purified and isolated human pluripotent granulocyte colony-stimulating factor free of association with any human protein in glycosylated or non-glycosylated form.
- 35. A pharmaceutical composition comprising an effective amount of a polypeptide according to claims 1 or 34 and a pharmaceutically acceptable diluent, adjuvant or carrier.
- 36. A method for providing hematopoietic therapy to a mammal comprising administering an effective amount of a polypeptide according to claims 1 or 34.
- 37. A method for arresting proliferation of leukemic cells comprising administering an effective amount of a polypeptide according to claims 1 or 34.
- 38. A DNA sequence coding for [Ser17]hpG-CSF.
- 39. A polypeptide product of the expression in a procaryotic or eucaryotic host cell of a DNA sequence according to claim 38.
- 40. A biologically functional plasmid or viral DNA vector including a DNA sequence according to claim 38.
- 41. A procaryotic or eucaryotic host cell stably transformed or transfected with a DNA vector according to claim 40.
- 42. A DNA sequence coding for an analog of hpG-CSF selected from the group consisting of:
- 43. A polypeptide product of the expression in a procaryotic or eucaryotic host cell of a DNA sequence according to claim 42.
- 44. A preparation of hpG-CSF which is greater than 95% pure and which comprises less than 0.5 ng of pyrogen per 0.5 mg of hpG-CSF.
Parent Case Info
[0001] This is a continuation-in-part of co-pending U.S. patent application Ser. No. 768,959, filed Aug. 23, 1985.
Divisions (4)
|
Number |
Date |
Country |
Parent |
09430103 |
Oct 1999 |
US |
Child |
09992873 |
Nov 2001 |
US |
Parent |
08679897 |
Jul 1996 |
US |
Child |
09060275 |
Apr 1998 |
US |
Parent |
08243556 |
May 1994 |
US |
Child |
08452135 |
May 1995 |
US |
Parent |
06835548 |
Mar 1986 |
US |
Child |
07269885 |
Nov 1988 |
US |
Continuations (4)
|
Number |
Date |
Country |
Parent |
09060275 |
Apr 1998 |
US |
Child |
09430103 |
Oct 1999 |
US |
Parent |
08452135 |
May 1995 |
US |
Child |
08679897 |
Jul 1996 |
US |
Parent |
08065465 |
May 1993 |
US |
Child |
08243556 |
May 1994 |
US |
Parent |
07269885 |
Nov 1988 |
US |
Child |
08065465 |
May 1993 |
US |
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
06768969 |
Aug 1985 |
US |
Child |
06835548 |
Mar 1986 |
US |