US4804744A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Osteogenic factors
Assignee: Biomet Manufacturing LLC
•
Filing: 1986-09-05
•
Granted: 1989-02-14
•
Expires: 2006-09-05
Abstract
Abstract A protein extracted from demineralized bone matrix exhibiting bone-inducing activity is disclosed as well as a method for the rapid isolation and purification of the protein to an essentially homogeneous state. Pharmaceutically acceptable compositions containing the bone-inducing protein and suitable pharmaceutical carriers are also described.
Inventors
- 👤 Arup Sen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 10:24
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/51; A; A61; A61K; A61K38/00; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/84
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
193 words
A method for isolating a preparation of an osteogenic protein which is a member of the P3 family of immunologically related proteins, said preparation characterized by being essentially homogeneous with respect to a molecular weight of from about 22,000 daltons to about 24,000 daltons from demineralized bone tissue comprising: (a) treating said demineralized bone tissue under aqueous conditions with a solubilizing agent for said osteogenic protein and thereby extracting the osteogenic factor into solution with said solubilizing agent; (b) subjecting said solution to size fractionation to recover a concentrated pool of proteins of molecular weight between about 12,000 daltons and 40,000 daltons; (c) subjecting said concentrated pool to a dialysis step comprising either (i) dialysis of said concentrated pool against an aqueous solvent containing trifluoroacetic acid and acetonitrile and removal of the insoluble material to obtain a soluble fraction or (ii) dialysis against deionized water to form an insoluble pellet and solubilization of said pellet with an aqueous solvent comprising trifluoroacetic acid and acetonitrile to obtain a soluble fraction; (d) subjecting the soluble fraction of step (c) to reverse phase high performance liquid chromatography; and (e) recovering a preparation of an osteogenic protein.
Claim 2
Independent Claim
193 words
A method for isolating a preparation of an osteogenic protein which is a member of the P3 family of immunologically related proteins, said preparation characterized by being essentially homogeneous with respect to a molecular weight of from about 22,000 daltons to about 24,000 daltons from demineralized bone tissue comprising: (a) treating said demineralized bone tissue under aqueous conditions with a solubilizing agent for said osteogenic protein and thereby extracting the osteogenic factor into solution with said solubilizing agent; (b) subjecting said solution to size fractionation to recover a concentrated pool of proteins of molecular weight between about 12,000 daltons and 40,000 daltons; (c) subjecting said concentrated pool to a dialysis step comprising either (i) dialysis of said concentrated pool against an aqueous solvent containing trifluoroacetic acid and acetonitrile and removal of the insoluble material to obtain a soluble fraction or (ii) dialysis against deionized water to form an insoluble pellet and solubilization of said pellet with an aqueous solvent comprising trifluoroacetic acid and acetonitrile to obtain a soluble fraction; (d) subjecting the soluble fraction of step (c) to reverse phase high performance liquid chromatography; and (e) recovering a preparation of an osteogenic protein.
Claim 3
Dependent Claim
11 words
The method of claim 1 wherein said solubilizing agent is guanidinium-hydrochloride.
Claim 4
Dependent Claim
35 words
The method of claim 1 wherein the solution isolated in step (a) is subjected to size fractionation to recover a concentrated pool of proteins of molcular weight between about 16,000 daltons and about 25,000 daltons.
Claim 5
Dependent Claim
17 words
The method of claim 1 wherein the size fractionation of step (b) is conducted by gel filtration.
Claim 6
Dependent Claim
63 words
A preparation of an osteogenic protein which is a member of the P3 family of immunologically related proteins, said preparation characterized by being essentially homogeneous with respect to a molecular weight of from about 22,000 daltons to about 24,000 daltons and by the capacity, in such essentially homogeneous state, of promoting osteogenesis in a manual prepared according to the process of claim 1.
Claim 7
Dependent Claim
11 words
The preparation of claim 5 which is isolated from bovine bone.
Claim 8
Dependent Claim
11 words
The preparation of claim 5 which is isolated from human bone.
Claim 9
Dependent Claim
11 words
The preparation of claim 5 which is isolated from porcine bone.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-02-14 | 1992-08-15 | 1993-02-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-02-14 | 1996-08-15 | 1997-02-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-02-14 | 2000-08-15 | 2001-02-14 | $7700.00 | Window Closed / Lapsed |