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US4439358A
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Method of preparing alpha-1-proteinase inhibitor
Assignee: Bayer Corp
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Filing: 1983-01-27
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Granted: 1984-03-27
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Expires: 2003-01-27
Abstract
Abstract A method is disclosed for separating alpha-1-proteinase inhibitor from a blood plasma fraction particularly from the Cohn fractionation scheme. An aqueous solution of the blood plasma fraction is held at a pH of about 6.5-8.5, and a temperature of about 2 DEG -50 DEG C. for a period of about 0.2-24 hours and then mixed with a poly-condensed polyglycol. The mixture may be held at a temperature of about 2 DEG -10 DEG C. for a period of about 1-24 hours. Next, the pH of the mixture is adjusted to selectively precipitate unwanted proteins from the solution of alpha-1-proteinase inhibitor which is separated from the solution and purified further. The amount of poly-condensed polyglycol and the pH of the mixture are selected accordingly.
Inventors
- 👤 Michael H. Coan
- 👤 William J. Brockway
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 17:54
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/81; C07K14/8107; C07K14/811; C07K14/8121; C07K14/8125; A; A61; A61K; A61K38/00; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/829; Y10S530/83; Y10S530/831
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
102 words
A method for separating alpha-1-proteinase inhibitor from a blood plasma fraction containing the same, which comprises the steps of (a) holding an aqueous solution of the blood plasma fraction at a pH of about 6.5-8.5, and a temperature of about 2°-50° C. for a period of about 0.2-24 hours, (b) mixing the solution with a polycondensed polyglycol and adjusting the pH of the mixture to selectively precipitate unwanted proteins from the solution without precipitation of alpha-1-proteinase inhibitor, the amount of polycondensed polyglycol and the pH of the mixture being determined according to FIG. 1, and (c) separating alpha-1-proteinase inhibitor from the solution.
Claim 2
Independent Claim
102 words
A method for separating alpha-1-proteinase inhibitor from a blood plasma fraction containing the same, which comprises the steps of (a) holding an aqueous solution of the blood plasma fraction at a pH of about 6.5-8.5, and a temperature of about 2°-50° C. for a period of about 0.2-24 hours, (b) mixing the solution with a polycondensed polyglycol and adjusting the pH of the mixture to selectively precipitate unwanted proteins from the solution without precipitation of alpha-1-proteinase inhibitor, the amount of polycondensed polyglycol and the pH of the mixture being determined according to FIG. 1, and (c) separating alpha-1-proteinase inhibitor from the solution.
Claim 3
Dependent Claim
19 words
The method of claim 1 wherein 1 part of polyglycol is used per 0.5-1.0 parts of blood plasma fraction.
Claim 4
Dependent Claim
40 words
The method of claim 1 wherein the mixture of the solution with the polyglycol is held at a temperature of about 2°-10° C. for a period of about 1-24 hours prior to the adjustment of the pH of the mixture.
Claim 5
Dependent Claim
20 words
The method of claim 1 which further includes the step of treating the alpha-1-proteinase inhibitor to render it hepatitis non-infective.
Claim 6
Dependent Claim
47 words
The method of claim 4 wherein the alpha-1-proteinase inhibitor is rendered hepatitis non-infective by heating an aqueous solution of the alpha-1-proteinase inhibitor in the presence of a heat stabilizing agent for a time and at a temperature and pH sufficient to render the alpha-1-proteinase inhibitor hepatitis non-infective.
Claim 7
Dependent Claim
16 words
The method of claim 5 wherein the heat stabilizing agent is a source of citrate ions.
Claim 8
Dependent Claim
13 words
The method of claim 5 wherein the heat stabilizing agent is a carbohydrate.
Claim 9
Dependent Claim
22 words
The method of claim 5 wherein the heat stabilizing agent is a mixture of a carbohydrate and a source of citrate ions.
Claim 10
Dependent Claim
31 words
The method of claim 5 wherein the alpha-1-proteinase inhibitor is heated for about 60° C. or more for a period up to about 10 hours at a pH of about 6.0-7.0.
Claim 11
Dependent Claim
25 words
The method of claim 1 wherein an effluent solution is separated from a precipitate in Step b prior to adjusting the pH of the mixture.
Claim 12
Dependent Claim
58 words
The method of claim 1 wherein the alpha-1-proteinase inhibitor is separated from the solution in step (c) by (d) separating a supernatant solution containing alpha-1-proteinase inhibitor from a precipitate, (e) contacting the solution with an anion exchange medium at a pH of about 5.5-8.6, and (f) selectively separating an effluent containing alpha-1-proteinase inhibitor from the anion exchange medium.
Claim 13
Dependent Claim
18 words
The method of claim 11 wherein the effluent of step (f) is treated to reduce its water content.
Claim 14
Dependent Claim
29 words
The method of claim 1 wherein the blood plasma fraction containing alpha-1-proteinase inhibitor is selected from the group consisting of Cohn Fraction IV-1, Cohn Fraction IV, and their reworks.
Claim 15
Dependent Claim
35 words
The method of claim 1 wherein the alpha-1-proteinase inhibitor is separated from the mixture in step (c) by contacting the mixture with a polycondensed polyglycol and separating an effluent from a precipitate containing alpha-1-proteinase inhibitor.
Claim 16
Dependent Claim
12 words
The method of claim 1 wherein the polycondensed polyglycol is polyethylene glycol.
Claim 17
Dependent Claim
16 words
The method of claim 15 wherein the molecular weight of the polyethylene glycol is about 3000-4000.
Claim 18
Dependent Claim
20 words
The method of claim 1 wherein the blood plasma fraction is treated with fumed silica to remove lipids contained therein.
Claim 19
Independent Claim
21 words
A method for removing lipids from Fraction IV, Fraction IV-1, or their reworks which comprises contacting said fraction with fumed silica.
Claim 20
Independent Claim
21 words
A method for removing lipids from Fraction IV, Fraction IV-1, or their reworks which comprises contacting said fraction with fumed silica.
Claim 21
Independent Claim
115 words
A method for separating alpha-1-proteinase inhibitor from a blood plasma fraction containing the same, which comprises the steps of (a) treating the blood plasma fraction containing alpha-1-proteinase to remove lipids contained therein, (b) holding an aqueous solution of the blood plasma fraction at a pH of about 6.5-8.5, and a temperature of about 2°-50° C. for a period of about 0.2-24 hours, (c) mixing the solution with a polycondensed polyglycol and adjusting the pH of the mixture to selectively precipitate unwanted proteins from the solution without precipitation of alpha-1-proteinase inhibitor, the amount of polycondensed polyglycol and the pH of the mixture being determined according to FIG. 1, and (d) separating alpha-1-proteinase inhibitor from the solution.
Claim 22
Independent Claim
115 words
A method for separating alpha-1-proteinase inhibitor from a blood plasma fraction containing the same, which comprises the steps of (a) treating the blood plasma fraction containing alpha-1-proteinase to remove lipids contained therein, (b) holding an aqueous solution of the blood plasma fraction at a pH of about 6.5-8.5, and a temperature of about 2°-50° C. for a period of about 0.2-24 hours, (c) mixing the solution with a polycondensed polyglycol and adjusting the pH of the mixture to selectively precipitate unwanted proteins from the solution without precipitation of alpha-1-proteinase inhibitor, the amount of polycondensed polyglycol and the pH of the mixture being determined according to FIG. 1, and (d) separating alpha-1-proteinase inhibitor from the solution.
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) | 1987-03-27 | 1987-09-26 | 1988-03-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-03-27 | 1991-09-26 | 1992-03-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-03-27 | 1995-09-26 | 1996-03-27 | $7700.00 | Window Closed / Lapsed |