Methods and compositions of human recombinant growth and differentiaton factor-5 (rhgdf-5) isolated from inclusion bodies
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 isolating recombinant GDF-5 protein from an inclusion body of a cell are disclosed herein. The methods as disclosed are cost-effective, time-saving and are of manufacturing quality.
Claims20
A method of isolating recombinant GDF-5 protein from an inclusion body of a cell, the method comprising:
- disrupting the cell such that a pGDF-5 or a pGDF-5-Trc-transformed cell releases the inclusion body of rhGDF-5 protein and recovering the GDF-5 protein from the inclusion body 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 substrate.
A method of isolating recombinant GDF-5 protein from an inclusion body of a prokaryotic cell, the method comprising:
- disrupting the prokaryotic cell via lysis or homogenization such that a pGDF-5 or a pGDF-5-Trc-transformed cell releases the inclusion body of rhGDF-5 protein and recovering the GDF-5 protein from the inclusion body 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 substrate.
A host cell line, the cell line comprising an inclusion body comprising an isolated GDF-5 protein, wherein the host cell includes an expression vector which comprises a polynucleotide encoding for an rhGDF-5 protein under the control of a T5 or Trc promoter.
Dependencies are inferred from the claim text.