Methods of purifying human recombinant growth and differentiation factor-5 (rhGDF-5) protein
AssigneeWarsaw Orthopedic, Inc.
InventorsDavid S. Scher, Roger E. Harrington
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Abstract
Expression vector systems are provided for increased production of a recombinant GDF-5 (rhGDF-5) protein. Also provided are transformed host cells that were engineered to produce and express high levels of rhGDF-5 protein. Methods for production and high expression of rhGDF-5 protein are disclosed herein. The methods of enhancing production and protein expression of rhGDF-5 protein as disclosed are cost-effective, time-saving and are of manufacturing quality.
Claims11
A method of purifying recombinant GDF-5 protein from a cell, the method comprising:
- providing a comprising an expression vector having a T5 or a Trc promoter operatively linked to a polynucleotide sequence that encodes a GDF-5 protein, and
- recovering the GDF-5 protein from the cell by contacting the GDF-5 protein with a substrate so as to isolate the GDF-5 protein,
- wherein the vector is a plasmid vector that is pGDF5-Trc or pGDF5-T5, the T5 or Trc promoter is inducible by isopropylthio-β-galactoside (IPTG), the pGDF5-T5 comprises the polynucleotide sequence of SEQ ID NO:
- 2 and the pGDF5-Trc comprises the polynucleotide sequence of SEQ ID NO:
- 3.
A method of purifying recombinant GDF-5 protein from a prokaryotic cell, the method comprising:
- recovering a GDF-5 protein from inclusion bodies of the prokaryotic cell, the prokaryotic cell comprising an expression vector having a T5 or a Trc promoter operatively linked to a polynucleotide sequence that encodes a GDF-5 protein by contacting the GDF-5 protein with a solid substrate so as to isolate the GDF-5 protein, wherein the vector is a plasmid vector that is pGDF5-Trc or pGDF5-T5, and the T5 or Trc promoter is inducible by isopropylthio-β-galactoside (IPTG), wherein the pGDF5-T5 comprises the polynucleotide sequence of SEQ ID NO:
- 2 and the pGDF5-Trc comprises the polynucleotide sequence of SEQ ID NO:
- 3.
Dependencies are inferred from the claim text.