US4434094A
Expired / Lapsed
US
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Partially purified osteogenic factor and process for preparing same from demineralized bone
Assignee: Celtrix Pharmaceuticals Inc
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Filing: 1983-04-12
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Granted: 1984-02-28
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Expires: 2003-04-12
Abstract
Abstract A process for partially purifying an osteogenic factor from demineralized bone particles in which nonfibrous proteins are extracted from the demineralized bone with a dissociative extractant for nonfibrous proteins, such as urea or guanidine hydrochloride, the extracted nonfibrous proteins are fractionated by anion exchange chromatography using DEAE cellulose at pH 7, the fraction not adsorbed by the DEAE cellulose is further fractionated by cation exchange chromatography using CM cellulose at pH 4.8, the fraction adsorbed by the CM cellulose is eluted therefrom and the partially purified osteogenic factor is recovered from the eluate as a ≦30,000 dalton protein isolate.
Inventors
- 👤 Saeid Seyedin
- 👤 Thomas Thomas
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 01:45
CPC Classifications
A; A61; A61L; A61L27/00; A61L27/14; A61L27/22; A61L27/227; A61F; A61F2/00; A61F2/02; A61F2/28; A61K; A61K35/00; A61K35/12; A61K35/32; A61L27/36; C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/51; A61K38/00; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/84; Y10S623/00; Y10S623/915; Y10S623/919
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
95 words
A process for partially purifying an osteogenic factor from particulate demineralized bone comprising: (a) extracting nonfibrous proteins from the demineralized bone with a liquid dissociative nonfibrous protein extractant; (b) contacting the extract of (a) with an anion exchanger at a pH of about 6.8 to about 7.2. (c) contacting the unadsorbed fraction of the extract with a cation exchanger at a pH of about 4.5 to about 5.2; (d) eluting the adsorbed fraction from the cation exchanger; and (e) isolating nonfibrous protein having a molecular weight below about 30,000 daltons from the eluate of (d).
Claim 2
Dependent Claim
95 words
A process for partially purifying an osteogenic factor from particulate demineralized bone comprising: (a) extracting nonfibrous proteins from the demineralized bone with a liquid dissociative nonfibrous protein extractant; (b) contacting the extract of (a) with an anion exchanger at a pH of about 6.8 to about 7.2. (c) contacting the unadsorbed fraction of the extract with a cation exchanger at a pH of about 4.5 to about 5.2; (d) eluting the adsorbed fraction from the cation exchanger; and (e) isolating nonfibrous protein having a molecular weight below about 30,000 daltons from the eluate of (d).
Claim 3
Dependent Claim
21 words
The process of claim 1 wherein steps (a) through (d) are each carried out in the presence of a protease inhibitor.
Claim 4
Dependent Claim
13 words
The process of claim 1 wherein the extractant is urea or guanidine hydrochloride.
Claim 5
Dependent Claim
13 words
The process of claim 1 wherein the extractant is 4 M guanidine hydrochloride.
Claim 6
Dependent Claim
17 words
The process of claim 1 wherein the anion exchanger and the cation exchanger are cellulosic ion exchangers.
Claim 7
Dependent Claim
42 words
The process of claim 5 wherein the anion exchanger is diethylaminoethyl cellulose and the cation exchanger is carboxymethyl cellulose; step (b) is carried out at a pH of about 7.0; and step (c) is carried out at a pH of about 4.8.
Claim 8
Dependent Claim
14 words
The process of claim 1 wherein step (e) is carried out by gel filtration.
Claim 9
Dependent Claim
66 words
The process of claim 2 wherein the extractant is 4 M guanidine, the extractant is removed from the extract, the nonfibrous proteins in the extract are dissolved in 6 M urea for contacting with the anion exchanger, the anion exchanger is diethylaminoethyl cellulose, the pH of step (b) is about 7.0, the cation exchanger is carboxymethyl cellulose, and the pH of step (c) is about 4.8.
Claim 10
Dependent Claim
11 words
Partially purified osteogenic factor prepared by the process of claim 1.
Claim 11
Dependent Claim
11 words
Partially purified osteogenic factor prepared by the process of claim 8.
References Cited (Backward Citations)
US3539549A
Cited
US4294753A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-02-27 | 1987-08-29 | 1988-02-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-02-27 | 1991-08-29 | 1992-02-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-02-27 | 1995-08-29 | 1996-02-28 | $7700.00 | Window Closed / Lapsed |