Patent US4720385A successfully refreshed from Google Patents.
US4720385A
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Protein compositions substantially free from infectious agents
Assignee: Bayer Corp
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Filing: 1985-05-20
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Granted: 1988-01-19
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Expires: 2005-05-20
Abstract
Abstract Compositions containing therapeutically or immunologically active proteins are rendered substantially free from infectious agents such as viable viruses and bacteria without substantial loss of therapeutic or immunologic activity by mixing the protein composition with a complex formed from transition metal ions, such as copper ions, and an angularly-fused, polynuclear heterocyclic arene having two nitrogen atoms in a "cis-ortho" relationship, such as phenanthroline, and a reducing agent such as a thiol or ascorbic acid or ascorbate salt or mixtures of ascorbic acid or ascorbate with a thiol in amounts and at a temperature and for a time sufficient to inactivate substantially all of the viruses and bacteria contained therein. Compositions containing therapeutically active proteins substantially free from viral and bacterial infectivity, which have heretofore been unattainable, can be prepared by the method of the invention.
Inventors
- 👤 Kenneth J. Lembach
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 03:09
CPC Classifications
A; A61; A61L; A61L2/00; A61L2/16; A61L2/18; A61L2/02; A61L2/04; A61L2103/00; A61L2103/05
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
226 words
A method for rendering a composition, which contains a therapeutically or immunologically active protein, selected from the group consisting of blood plasma proteins, virus vaccines, bacterial vaccines, and non-infectious antigens dissolved in an aqueous medium, substantially free from infectious agents selected from viable viruses and bacteria without substantial loss of therapeutic or immunologic activity, which comprises (a) mixing an aqueous solution of the protein composition with an effective amount of a complex formed from mixing an angularly-fused, polynuclear heterocyclic arene having two nitrogen atoms in a "cis-ortho" relationship and a transition metal ion capable of complexing with said arene and a reducing agent, selected from ascorbic acid, ascorbate group present as an ester or as the anion component of an alkali metal or alkaline earth metal salt of ascorbic acid, a mixture of ascorbic acid or ascorbate with a thiol, NADPH, NADH, and a mixture of NADPH or NADH with at least one of a thiol, ascorbic acid and ascorbate in an effective amount, said amounts being effective to inactivate substantially all of the infectious agents contained in said composition and which does not result in substantial loss of thereapeutic or immunologic activity, and (b) holding the mixture for a time and at a temperature sufficient to render said infectious agents substantially non-infective in the protein composition without substantial loss of therapeutic or immunologic activity therein.
Claim 2
Independent Claim
226 words
A method for rendering a composition, which contains a therapeutically or immunologically active protein, selected from the group consisting of blood plasma proteins, virus vaccines, bacterial vaccines, and non-infectious antigens dissolved in an aqueous medium, substantially free from infectious agents selected from viable viruses and bacteria without substantial loss of therapeutic or immunologic activity, which comprises (a) mixing an aqueous solution of the protein composition with an effective amount of a complex formed from mixing an angularly-fused, polynuclear heterocyclic arene having two nitrogen atoms in a "cis-ortho" relationship and a transition metal ion capable of complexing with said arene and a reducing agent, selected from ascorbic acid, ascorbate group present as an ester or as the anion component of an alkali metal or alkaline earth metal salt of ascorbic acid, a mixture of ascorbic acid or ascorbate with a thiol, NADPH, NADH, and a mixture of NADPH or NADH with at least one of a thiol, ascorbic acid and ascorbate in an effective amount, said amounts being effective to inactivate substantially all of the infectious agents contained in said composition and which does not result in substantial loss of thereapeutic or immunologic activity, and (b) holding the mixture for a time and at a temperature sufficient to render said infectious agents substantially non-infective in the protein composition without substantial loss of therapeutic or immunologic activity therein.
Claim 3
Dependent Claim
29 words
The method of claim 1 wherein the mixture is held in step (b) at a temperature of about 2°-60° C. for a period of at least about 1 minute.
Claim 4
Dependent Claim
29 words
The method of claim 1 wherein the mixture is held in step (b) at a temperature of about 20°-37° C. for a period of at least about 0.25 hours.
Claim 5
Dependent Claim
32 words
The method of claim 1 wherein the mixture is held for a time and at a temperature to inactivate at least about 99% of the viruses and bacteria in the protein composition.
Claim 6
Dependent Claim
14 words
The method of claim 1 wherein the pH in step (b) is physiologically compatible.
Claim 7
Dependent Claim
11 words
The method of claim 1 wherein the pH is about 4-10.
Claim 8
Dependent Claim
29 words
The method of claim 1 wherein the amount of complex in step (a) is about 0.0001-0.1 mM in an aqueous solution containing about 0.005-10% (weight/volume) of the protein composition.
Claim 9
Dependent Claim
36 words
The method of claim 1 wherein the amount of reducing agent is at least about 1 mM in the absence of added hydrogen peroxide in an aqueous solution containing about 0.005-10% (weight/volume) of the protein composition.
Claim 10
Dependent Claim
12 words
The method of claim 1 wherein the complex is a copper/1,10-phenanthroline complex.
Claim 11
Dependent Claim
67 words
The method of claim 1 wherein the reducing agent is selected from (a) ascorbic acid, (b) ascorbate group present as the anion component of one of an alkali metal and an alkaline earth metal salt, and (c) a mixture of one of ascorbic acid and ascorbate group present as the anion component of one of an alkali metal and an alkaline earth metal salt with a thiol.
Claim 12
Dependent Claim
40 words
The method of claim 1 wherein the reducing agent is a mixture of one of ascorbic acid and ascorbate group present as the anion component of one of an alkali metal and an alkaline earth metal salt with a thiol.
Claim 13
Dependent Claim
24 words
The method of claim 1 wherein the reducing agent is a mixture of one of cysteine and 3-mercaptopropionic acid with ascorbic acid (sodium salt).
Claim 14
Dependent Claim
40 words
The method of claim 1 including the further step of: (c) removing the added complex formed from the angularly-fused, polynuclear heterocyclic arene and the transition metal ion, low molecular weight reaction components, and low molecular weight products of reaction components.
Claim 15
Dependent Claim
16 words
The method of claim 13 wherein the protein composition comprises a component derived from blood plasma.
Claim 16
Dependent Claim
13 words
The method of claim 13 wherein the protein composition comprises a viral vaccine.
Claim 17
Dependent Claim
12 words
The method of claim 13 wherein the protein composition comprises non-infectious antigen.
Claim 18
Dependent Claim
111 words
The method of claim 13 wherein the protein composition comprises a protein selected from the group consisting of blood plasma, partially fractionated blood plasma, plasminogen, albumin, antihemophilic factor (Factor VIII), Factor IX concentrate containing Factors II and VII and IX and X, Factor II, Factor VII, Factor IX, Factor X, Plasma Protein Fraction (human), fibronectin (cold insoluble globulin), Factor XIII, IgG, IgA, IgD, IgE, IgM, high molecular weight kininogen (90,000-106,000), an immune globulin, intravenous (modified, either chemically or enzymatically or by fractional separation) immune serum globulin, monoclonal antibodies, anti-inhibitor coagulant complex (FEIBA), antithrombin III, alpha-1-proteinase inhibitor, plasma growth hormone, somatomedin, prealbumin, plasminogen-streptokinase complex, ceruloplasmin, transferrin, haptoglobin, and prekallikrein and mixtures thereof.
Claim 19
Dependent Claim
9 words
A product produced by the method of claim 14.
Claim 20
Dependent Claim
9 words
A product produced by the method of claim 15.
Claim 21
Dependent Claim
9 words
A product produced by the method of claim 16.
Claim 22
Dependent Claim
9 words
A product produced by the method of claim 17.
Claim 23
Independent Claim
244 words
A method for rendering non-infective and innocuous, without substantial loss of activity, a composition which contains a protein produced by means of biotechnology by protein-producing organisms in a suitable medium which comprises (a) contacting said medium containing at least one of (i) intact protein-producing organisms, and (ii) adventitious agents such as viruses, and (iii) nucleic acid genetic material from the protein-producing organisms or adventitious agents with an aqueous solution of an effective amount of a complex formed from mixing an angularly-fused, polynuclear heterocyclic arene having two nitrogen atoms in a "cis-ortho" relationship and a transition metal ion capable of complexing with said arene and a reducing agent, selected from a thiol, ascorbic acid, ascorbate group present as an ester or as the anion component of an alkali metal or alkaline earth metal salt of ascorbic acid, a mixture of ascorbic acid or ascorbate with a thiol, NADPH, NADH, and a mixture of NADPH or NADH with at least one of a thiol, ascorbic acid and ascorbate in an effective amount, said amounts being effective to inactivate substantially all of the infectious agents contained in said composition and which does not result in substantial loss of activity of the protein, and (b) holding the mixture from Step (a) for a time and at a temperature sufficient to render said protein-producing organisms, adventitious agents, and nucleic acid genetic material from said protein-producing organisms and adventitious agents substantially non-infective and innocuous without substantial loss of protein activity.
Claim 24
Independent Claim
244 words
A method for rendering non-infective and innocuous, without substantial loss of activity, a composition which contains a protein produced by means of biotechnology by protein-producing organisms in a suitable medium which comprises (a) contacting said medium containing at least one of (i) intact protein-producing organisms, and (ii) adventitious agents such as viruses, and (iii) nucleic acid genetic material from the protein-producing organisms or adventitious agents with an aqueous solution of an effective amount of a complex formed from mixing an angularly-fused, polynuclear heterocyclic arene having two nitrogen atoms in a "cis-ortho" relationship and a transition metal ion capable of complexing with said arene and a reducing agent, selected from a thiol, ascorbic acid, ascorbate group present as an ester or as the anion component of an alkali metal or alkaline earth metal salt of ascorbic acid, a mixture of ascorbic acid or ascorbate with a thiol, NADPH, NADH, and a mixture of NADPH or NADH with at least one of a thiol, ascorbic acid and ascorbate in an effective amount, said amounts being effective to inactivate substantially all of the infectious agents contained in said composition and which does not result in substantial loss of activity of the protein, and (b) holding the mixture from Step (a) for a time and at a temperature sufficient to render said protein-producing organisms, adventitious agents, and nucleic acid genetic material from said protein-producing organisms and adventitious agents substantially non-infective and innocuous without substantial loss of protein activity.
Claim 25
Dependent Claim
40 words
The method of claim 22 including the further step of: (c) removing the added complex formed from the angularly-fused, polynuclear heterocyclic arene and the transition metal ion, low molecular weight reaction components, and low molecular weight products of reaction components.
Claim 26
Dependent Claim
9 words
A product produced by the method of claim 23.
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) | 1991-01-18 | 1991-07-20 | 1992-01-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-01-18 | 1995-07-20 | 1996-01-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-01-18 | 1999-07-20 | 2000-01-19 | $7700.00 | Window Closed / Lapsed |