US4780529A
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Isolation of an endotoxin inactivator from human plasma, and methods of use
Assignee: Cambridge Biotech Corp
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Filing: 1987-06-16
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Granted: 1988-10-25
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Expires: 2007-06-16
Abstract
Abstract An endotoxin inactivator is isolated from human plasma by anion exchange (DEAE-Sephadex), dye-affinity (Cibracron Blue-Sepharose), and adsorption (on hydroxyapatite) chromatography. The endotoxin inactivator, isolated in essentially pure form, may be used to depyrogenate clinical blood products.
Inventors
- 👤 Yu-Lee Hao
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 07:53
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4702; C07K14/4703; A; A61; A61L; A61L2/00; A61L2/16; A61L2/18; A61L2103/00; A61L2103/05; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/829
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
70 words
A method of depyrogenating a pyrogenic blood product which comprises providing a pyrogenic blood product, and adding an endotoxin inactivating amount of endotoxin inactivator derived from blood and characterized as follows: (a) a molecular weight of about 61,000 daltons; (b) elution of about 0.2-0.3M NaCl form a DEAE-Sephadex adsorbent; (c) elution at about 01.5-0.25M NaCl from a Cibacron-Blue-Sepharose adsorbent; and (d) elution at about 0.1-0.25M phosphate from a hydroxyapatite adsorbent.
Claim 2
Independent Claim
70 words
A method of depyrogenating a pyrogenic blood product which comprises providing a pyrogenic blood product, and adding an endotoxin inactivating amount of endotoxin inactivator derived from blood and characterized as follows: (a) a molecular weight of about 61,000 daltons; (b) elution of about 0.2-0.3M NaCl form a DEAE-Sephadex adsorbent; (c) elution at about 01.5-0.25M NaCl from a Cibacron-Blue-Sepharose adsorbent; and (d) elution at about 0.1-0.25M phosphate from a hydroxyapatite adsorbent.
Claim 3
Dependent Claim
15 words
The method of claim 1 wherein the endotoxin inactivator is provided in essentially pure form.
Claim 4
Dependent Claim
37 words
The method of claim 2, wherein the inactivator is capable of inactivating at least about: 3,657 ng S. typhosa endotoxin, 914 ng S. enteritides endotoxin, or 974 ng E. coli 055:B5 endotoxin, per milligram of endotoxin inactivator.
Claim 5
Dependent Claim
32 words
The method of claim 2, wherein the endotoxin inactivator is provided in essentially pure form by subjecting an endotoxin-inactivator-containing fraction of blood to DEAE-sephadex chromatography, immobilized chromophore blue chromatography, and hydroxyapatite chromatography.
Claim 6
Dependent Claim
13 words
The method of claim 2 in which the endotoxin inactivator is essentially stable.
Claim 7
Dependent Claim
25 words
An essentially nonpyrogenic blood product suitable for parenteral administration consisting essentially of a first blood protein and an essentially stable endotoxin inactivator derived from blood.
Claim 8
Dependent Claim
25 words
An essentially nonpyrogenic blood product suitable for parenteral administration consisting essentially of a first blood protein and an essentially stable endotoxin inactivator derived from blood.
Claim 9
Dependent Claim
14 words
The product of claim 6 in which the first blood protein is an immunoglobulin.
Claim 10
Dependent Claim
14 words
The product of claim 6 in which the first blood protein is antihemophilic factor.
Claim 11
Dependent Claim
14 words
The product of claim 6 in which the first blood protein is Factor IX.
Claim 12
Dependent Claim
14 words
The product of claim 6 in which the first blood protein is an interferon.
Claim 13
Independent Claim
96 words
A method for the purification of Factor IX complex and endotoxin inactivator from which comprises: (a) subjecting the plasma to DEAE-Sephadex chromatography; (b) eluting the endotoxin inactivator in partially purified from with an eluting buffer containing about 0.2-0.3M NaCl; (c) eluting the Factor IX complex with an eluting buffer containing about 2M NaCl; (d) subjecting the partially purified endotoxin inactivator of step (b) to Cibacron-blue Sepharose chromatography to yield a further purified endotoxin inactivator; and (e) subjecting the further purified endotoxin inactivator of step (d) to hydroxyapatite chromatography to yield endotoxin inactivator in essentially pure form.
Claim 14
Independent Claim
96 words
A method for the purification of Factor IX complex and endotoxin inactivator from which comprises: (a) subjecting the plasma to DEAE-Sephadex chromatography; (b) eluting the endotoxin inactivator in partially purified from with an eluting buffer containing about 0.2-0.3M NaCl; (c) eluting the Factor IX complex with an eluting buffer containing about 2M NaCl; (d) subjecting the partially purified endotoxin inactivator of step (b) to Cibacron-blue Sepharose chromatography to yield a further purified endotoxin inactivator; and (e) subjecting the further purified endotoxin inactivator of step (d) to hydroxyapatite chromatography to yield endotoxin inactivator in essentially pure form.
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) | 1991-10-25 | 1992-04-25 | 1992-10-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-10-25 | 1996-04-25 | 1996-10-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-10-25 | 2000-04-25 | 2000-10-25 | $7700.00 | Window Closed / Lapsed |