Expression conditions and methods of human recombinant growth and differentiation factor-5 (rhGDF-5)
US-9051389-B2 · Jun 9, 2015 · US
Host cell lines expressing recombinant GDF-5 protein
US9540429B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9540429-B2 |
| Application number | US-201514610590-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jan 30, 2015 |
| Priority date | Jan 25, 2013 |
| Publication date | Jan 10, 2017 |
| Grant date | Jan 10, 2017 |
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Abstract
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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.
First claim
Opening claim text (preview).
What is claimed is: 1. A host cell line, the cell line comprising an inclusion body comprising an isolated rhGDF-5 protein, wherein the host cell includes an expression vector comprising a polynucleotide encoding for an rhGDF-5 protein under the control of a T5 promoter, the rhGDF-5 protein comprising the amino acid sequence of SEQ ID NO: 4. 2. A method of producing an rhGDF-5 protein, the method comprising providing a host cell line having an expression vector comprising a polynucleotide encoding for the rhGDF-5 protein under the control of a T5 promoter; and recovering the rhGDF-5 protein from the host cell line, wherein the rhGDF-5 protein comprises the amino acid sequence of SEQ ID NO: 4. 3. A host cell line of claim 1 , wherein the host cell line is an Escherichia coli cell comprising the strain of DH10β, STBL2, HMS174 and RecA. 4. A host cell line of claim 1 , wherein the expression vector further comprises a Kanamycin-resistant gene (Kan r gene). 5. A method of claim 2 , wherein the rhGDF-5 protein is recovered and purified via ammonium sulfate or ethanol precipitation, acid or base extraction, column chromatography, affinity column chromatography, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, hydroxylapatite chromatography, lectin chromatography and/or gel electrophoresis. 6. A method of claim 2 , wherein the host cell line is an Escherichia coli cell comprising the strain of DH10β, STBL2, HMS174 and RecA. 7. A method of claim 2 , wherein the expression vector further comprises a Kanamycin-resistant gene (Kan r gene).
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Classifications
Extraction; Separation; Purification · CPC title
- C07K14/475Primary
Growth factors; Growth regulators · CPC title
- C07K14/51Primary
Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor · CPC title
by a combination of two or more processes of different types · CPC title
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- What does patent US9540429B2 cover?
- 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…
- Who is the assignee on this patent?
- Warsaw Orthopdedic Inc, Warsaw Orthopedic Inc
- What technology area does this patent fall under?
- Primary CPC classification C07K14/475. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Jan 10 2017 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 2 related publications on this page (citations in our corpus or others sharing the same primary CPC).