US5408039A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for an industrial-scale preparation of a standardized human von Willebrand factor concentrate of very high purity and suitable for therapeutic use
Assignee: L'ESTABLISSEMENT FRANCAIS DU SANG (FRENCH BLOOD BANK)
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Filing: 1992-03-06
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Granted: 1995-04-18
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Expires: 2012-03-06
Abstract
Abstract The invention relates to a process for purifying human von Willebrand factor from a cryoprecipitated plasma fraction, which comprises a combination of three chromatographic separation steps. The first chromatographic separation step comprises contacting a cryoprecipitated fraction with a large-pore vinyl polymer resin having DEAE group. The effluent from this separation step is again contacted with a large pore vinyl polymer resin having DEAE groups in the second chromatographic step. In the third chromatographic separation step, the effluent from the second step is subjected to affinity chromatography by contacting with gelatin-Sepharose. The concentrate obtained has very high specific activity and a high percentage of high molecular weight multimers. The concentrate is intended, in particular, for therapeutic use.
Inventors
- 👤 Miryana Burnouf-Radosevich
- 👤 Thierry Burnouf
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 20:46
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/745; C07K14/755; A; A61; A61P; A61P7/00; A61P7/04; A61K; A61K38/00
Claims Analysis
Total Claims: 8
Claim 1
Independent Claim
136 words
A process for preparing a purified concentrate of human von Willebrand factor enriched in high molecular weight multimers comprising: (a) subjecting a cryoprecipitated fraction of plasma to a first ion exchange chromatography step comprising contacting said cryoprecipitated fraction with a large-pore vinyl polymer resin having DEAE groups attached thereto, and obtaining a first von Willebrand factor-containing effluent; (b) subjecting said first von Willebrand factor-containing effluent to a second ion exchange chromatography step comprising contacting said first von Willebrand factor-containing effluent with a large-pore vinyl polymer resin having DEAE groups attached thereto and obtaining a second von Willebrand factor-containing effluent; and (c) subjecting said second von Willebrand factor-containing effluent to an affinity chromatography step comprising contacting with gelatin-SEPHAROSE, and obtaining an effluent containing a purified concentrate of human von Willebrand factor enriched in high molecular weight multimers.
Claim 2
Dependent Claim
136 words
A process for preparing a purified concentrate of human von Willebrand factor enriched in high molecular weight multimers comprising: (a) subjecting a cryoprecipitated fraction of plasma to a first ion exchange chromatography step comprising contacting said cryoprecipitated fraction with a large-pore vinyl polymer resin having DEAE groups attached thereto, and obtaining a first von Willebrand factor-containing effluent; (b) subjecting said first von Willebrand factor-containing effluent to a second ion exchange chromatography step comprising contacting said first von Willebrand factor-containing effluent with a large-pore vinyl polymer resin having DEAE groups attached thereto and obtaining a second von Willebrand factor-containing effluent; and (c) subjecting said second von Willebrand factor-containing effluent to an affinity chromatography step comprising contacting with gelatin-SEPHAROSE, and obtaining an effluent containing a purified concentrate of human von Willebrand factor enriched in high molecular weight multimers.
Claim 3
Dependent Claim
20 words
The process according to claim 1, wherein said cryoprecipitated fraction of plasma has been prepurified by contact with aluminum hydroxide.
Claim 4
Dependent Claim
46 words
The process according to claim 1, wherein said large-pore vinyl polymer resin having DEAE groups attached thereto in steps (a) and (b) is DEAE-Fractogel® TSK 650 equilibrated with a buffer solution containing 0.01M trisodium citrate, 0.11M sodium chloride, 0.001M calcium chloride, 0.12M glycine, and 0.016M L-lysine.
Claim 5
Dependent Claim
57 words
The process according to claim 1, wherein said first ion exchange chromatography step (a) comprises introducing said cryoprecipitated fraction of plasma to a first ion exchange chromatography column, which has been equilibrated with a buffer solution, and eluting said first von Willebrand factor containing effluent by increasing the sodium chloride concentration of said buffer solution to 0.14-0.15M.
Claim 6
Dependent Claim
57 words
The process according to claim 1, wherein said second ion exchange chromatography step (b) comprises introducing said cryoprecipitated fraction of plasma to a second ion exchange chromatography column, which has been equilibrated with a buffer solution, and eluting said second von Willebrand factor containing effluent by increasing the sodium chloride concentration of said buffer solution to 0.15-0.17M.
Claim 7
Dependent Claim
66 words
The process according to claim 1, wherein said affinity chromatography step (c) comprises selectively adsorbing residual fibronectin by introducing the effluent from said second ion exchange chromatography step (b) to a gelatin-SEPHAROSE chromatography column equilibrated with the elution buffer from said second ion exchange chromatography step (b) and obtaining an effluent containing a purified concentrate of human von Willebrand factor enriched in high molecular weight multimers.
Claim 8
Dependent Claim
31 words
The process according to claim 1, wherein said obtaining of said purified concentrate comprises collecting, dispensing, and freeze-drying the von Willebrand factor present in the effluent of said affinity chromatography step.
References Cited (Backward Citations)
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1998-04-17 | 1998-10-17 | 1999-04-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2002-04-17 | 2002-10-17 | 2003-04-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2006-04-17 | 2006-10-17 | 2007-04-18 | $7700.00 | Window Closed / Lapsed |