Glial derived neurotrophic factor
AssigneeAmgen Inc.
InventorsLeu-Fen H. Lin, Franklin D. Collins, Daniel H. Doherty, Jack Lile, Susan Bektesh
Life of this patent
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Abstract
A novel neurotrophic factor referred to as glial derived neurotrophic factor (GDNF) has been identified and isolated from serum free growth conditioned medium of B49 glioblastoma cells. Rat and human genes encoding GDNF have been cloned and sequenced. A gene encoding GDNF has been subcloned into a vector, and the vector has been used to transform a host cell in order to produce biologically active GDNF in a recombinant DNA process.
Claims74
Substantially purified glial derived neurotrophic factor.
A substantially purified glial derived neurotrophic factor characterized by the following:
- (a) an apparent molecular weight of about 31-42 kD on non-reducing SDS-PAGE;
- (b) an apparent molecular weight of about 20-23 kD on reducing SDS-PAGE;
- and
- (c) a specific activity of at least about 12,000 TU/mg.
A pharmaceutical composition comprising an effective amount of substantially purified glial derived neurotrophic factor in a pharmaceutically suitable carrier.
A method for obtaining substantially purified glial derived neurotrophic factor comprising:
- preparing a serum-free growth-conditioned medium of B49 glioblastoma cells;
- concentrating the conditioned medium;
- performing heparin sepharose chromatography on the concentrated conditioned medium;
- performing fast protein liquid chromatography or fractions obtained from said heparin sepharose chromatography;
- and
- performing reverse-phase high-performance liquid chromatography or fractions obtained from said fast protein liquid chromatography.
A purified and isolated nucleic acid sequence encoding glial derived neurotrophic factor.
A purified and isolated nucleic acid sequence encoding pre-pro glial derived neurotrophic factor.
A method for preventing or treating nerve damage which comprises administering to a patient in need thereof a therapeutically effective amount of glial derived neurotrophic factor.
A pharmaceutical composition for preventing or treating Parkinson's disease comprising an effective amount of glial derived neurotrophic factor in a pharmaceutically suitable carrier.
A pharmaceutical composition for preventing or treating damaged or improperly functioning dopaminergic nerve cells comprising an effective amount of glial derived neurotrophic factor in a pharmaceutically suitable carrier.
A method for preventing damage to or treating damaged or improperly functioning dopaminergic nerve cells which comprises administering to a patient in need thereof a therapeutically effective amount of glial derived neurotrophic factor.
A recombinant DNA molecule comprising expression regulatory elements operatively linked to a nucleic acid sequence encoding glial derived neurotrophic factor.
A recombinant DNA method for production of glial derived neurotrophic factor comprising:
- (a) subcloning a DNA sequence encoding for glial derived neurotrophic factor into an expression vector which comprises the regulatory elements needed to express the DNA sequence;
- (b) transforming a host cell with said expression vector;
- (c) culturing the host cells under conditions for amplification of the vector and expression of glial derived neurotrophic factor;
- and
- (d) harvesting the glial derived neurotrophic factor from the host cell culture.
Substantially purified antibodies that recognize glial derived neurotrophic factor.
A method for preventing or treating nerve damage which comprises implanting cells that secrete glial derived neurotrophic factor into the body of patients in need thereof.
A device for preventing or treating nerve damage by implantation in a patient comprising:
- a semipermeable membrane;
- and
- a cell that secretes glial derived neurotrophic factor encapsulated within said membrane,
- said membrane being permeable to said glial derived neurotrophic factor, and impermeable to factors from said patents detrimental to said cells.
Dependencies are inferred from the claim text.