EP0121976A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Partially purified osteogenic factor and process for preparing same from demineralized bone or an osteosarcoma
Assignee: Collagen Aesthetics Inc
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Filing: 1984-01-19
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Granted: —
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Expires: 2004-01-19
Abstract
Abstract A process for partially purifying an osteogenic factor from demineralized bone particles or osteosarcomas in which nonfibrous proteins are extracted from the demineralized bone or osteosarcomas with a dissociative extractant for nonfibrous proteins, such as urea or guanl- dine 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
Inventors
- 👤 Saeid Seyedin
- 👤 Thomas Thomas
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 15:46
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: 10
Claim 1
Independent Claim
116 words
A process for partially purifying an osteogenic factor from particulate demineralized bone or an osteosarcoma comprising extracting nonfibrous proteins from the demineralized bone or osteosarcoma with a liquid dissociative nonfibrous protein extractant and separating the osteogenic factor from the extract by ion exchange chromatography characterized in that: (a) the extract is first contacted with an anion exchanger at a pH of about 6.8 to about 7.2; (b) the unadsorbed fraction from the anion exchanger is contacted with a cation exchanger at a pH of about 4.5 to about 5.2; (c) the adsorbed fraction from the cation exchanger is eluted; and d) nonfibrous protein having a molecular weight below about 30,000 daltons are isolated from the eluate.
Claim 2
Dependent Claim
116 words
A process for partially purifying an osteogenic factor from particulate demineralized bone or an osteosarcoma comprising extracting nonfibrous proteins from the demineralized bone or osteosarcoma with a liquid dissociative nonfibrous protein extractant and separating the osteogenic factor from the extract by ion exchange chromatography characterized in that: (a) the extract is first contacted with an anion exchanger at a pH of about 6.8 to about 7.2; (b) the unadsorbed fraction from the anion exchanger is contacted with a cation exchanger at a pH of about 4.5 to about 5.2; (c) the adsorbed fraction from the cation exchanger is eluted; and d) nonfibrous protein having a molecular weight below about 30,000 daltons are isolated from the eluate.
Claim 3
Dependent Claim
24 words
The process of claim 1 further characterized in that steps (a) through (c) are each carried out in the presence of a protease inhibitor.
Claim 4
Dependent Claim
21 words
The process of claim 1 further . characterized in that the anion exchanger and the cation exchanger are cellulosic ion exchangers.
Claim 5
Dependent Claim
45 words
The process of claim 3 further characterized in that the anion exchanger is diethylaminoethyl cellulose and the cation exchanger is carboxymethyl cellulose; step (a) is carried out at a pH of about 7.0; and step (b) is carried out at a pH of about 4.8.
Claim 6
Dependent Claim
17 words
The process of claim 1 further characterized in that step (d) is carried out by gel filtration.
Claim 7
Dependent Claim
63 words
The process of claim 2 further characterized in that the extractant is removed from the extract, the nonfibrous proteins in the extracts are dissolved in 6 mM urea for contacting with the anion exchanger, the anion exchanger is diethylaminoethyl cellulose, the pH of step (a) is about 7.0, and cation exchanger is carboxymethyl cellulose, and the pH of step (c) is about 4.8.
Claim 8
Dependent Claim
11 words
Partially purified osteogenic factor prepared by the process of claim 1.
Claim 9
Dependent Claim
53 words
Partially purified osteogenic factor present in vertebrate bone characterized in that it: (a) is nonfibrous; (b) is unadsorbed by diethylaminoethyl cellulose anion exchanger at a pH of about 7.0; (c) is adsorbed by carboxymethyl cellulose cation exchanger at a pH of about 4.8; and (d) has a molecular weight below about 30,000 daltons.
Claim 10
Dependent Claim
53 words
Partially purified osteogenic factor present in vertebrate bone characterized in that it: (a) is nonfibrous; (b) is unadsorbed by diethylaminoethyl cellulose anion exchanger at a pH of about 7.0; (c) is adsorbed by carboxymethyl cellulose cation exchanger at a pH of about 4.8; and (d) has a molecular weight below about 30,000 daltons.
References Cited (Backward Citations)
No citation records indexed.
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