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US4341867A Expired / Lapsed US ⏱️ 0.0 years remaining

Process for recovering enzymes from blood

Assignee: DE FORENDE BRYGGERIER AS
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Filing: 1980-05-13
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Granted: 1982-07-27
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Expires: 2000-05-13
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Abstract

Abstract Various enzymes, in particular Cu,Zn-superoxide dismutase (SOD), catalase and carbonic acid anhydrase, are recovered from blood by admixing wholly or partly isolated blood cells with ethanol or a homologous alcohol until a concentration of 10 to 70% by volume and allowing them to stand for hemolysis of the blood cells and denaturation of the hemoglobin, and then adding water up to the double volume or more, and removing the precipitate of cell residuals, hemoglobin and other denatured proteins from the suspension. Then the desired enzymes are isolated from the solution obtained. In particular, SOD and catalase can both be isolated by chromatography of the solution at a pH of 4.7 to 5.5 on a cation exchange resin of the same polarity as SOD in the pH range used and elution of the resin with a buffer solution which has a pH in the range 4.7 to 7.5 and an ionic strength in the range 0.01 to 1.0 M, SOD being eluted at the lowest pH and/or the lowest ionic strength. The process allows both the hemolysis of the blood cells and the precipitation of the hemoglobin to be carried out in one step while retaining the activity of the desired enzymes, so that they can be obtained in a high yield in a manner which lends itself to use on an industrial scale. Further, SOD and catalase may be isolated and purified in a single chromatography step.

Inventors

  • 👤 Jack T. Johansen

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-06 17:44
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0089; C12N9/0065; C12N9/88; Y; Y10; Y10S; Y10S435/00; Y10S435/814; Y10S435/816

Claims Analysis

Total Claims: 7
Claim 1
Independent Claim 120 words
A process for recovery one or more of the enzymes Cu,Zn-superoxide dismutase (SOD), catalase and carbonic acid anhydrase from blood by lysing the blood cells and isolating the desired enzymes from the solution obtained, characterized by admixing wholly or partly isolated blood cells with an alkanol having 2 to 4 carbon atoms until a concentration of 10 to 70% by volume and allowing them to stand for hemolysis of the blood cells and denaturation of the hemoglobin and then adding water in an amount so as at least approximately to double the volume of the mixture, removing the precipitate of cell residuals, hemoglobin and other denatured proteins from the suspension, and finally isolating the desired enzymes from the solution obtained.
Claim 2
Dependent Claim 120 words
A process for recovery one or more of the enzymes Cu,Zn-superoxide dismutase (SOD), catalase and carbonic acid anhydrase from blood by lysing the blood cells and isolating the desired enzymes from the solution obtained, characterized by admixing wholly or partly isolated blood cells with an alkanol having 2 to 4 carbon atoms until a concentration of 10 to 70% by volume and allowing them to stand for hemolysis of the blood cells and denaturation of the hemoglobin and then adding water in an amount so as at least approximately to double the volume of the mixture, removing the precipitate of cell residuals, hemoglobin and other denatured proteins from the suspension, and finally isolating the desired enzymes from the solution obtained.
Claim 3
Dependent Claim 26 words
A process according to claim 1, characteristized by isolating one or more of the enzymes Cu,Zn-superoxide dismutase (SOD), catalase and carbonic acid anhydrase from the solution.
Claim 4
Dependent Claim 90 words
A process according to claim 2, characterized by the fact that SOD and catalase are isolated from the solution by subjecting said solution at a pH of about 4.8 to chromatography on a cation exchange resin of the same polarity as SOD in the pH range used, eluting SOD and catalase from the resin with a buffer solution which has a pH in the range 4.7 to 7.5 and an ionic strength in the range 0.01 to 1.0 M, SOD being eluted at the lowest pH and/or lowest ionic strength.
Claim 5
Dependent Claim 59 words
A process according to claim 3, characterized by eluting SOD from the cation exchange resin with a buffer solution having a pH of 4.7 to 5.5 and an ionic strength of 0.02 to 0.2 M and eluting the catalase with a buffer solution having a pH of 6.5 to 7.5 and an ionic strength of 0.1 to 1.0 M.
Claim 6
Dependent Claim 34 words
A process according to claim 3 or 4, characterized by using as cation exchange resin carboxymethyl celluloses or cross-linked dextrans substituted with carboxymethyl groups or sulfopropyl groups or cross-linked agaroses substituted with carboxymethyl groups.
Claim 7
Dependent Claim 18 words
A process according to any one of claims 1, 2, 3 and 4 wherein said alkanol is ethanol.

References Cited (Backward Citations)

GB720232A Cited
CH478097A Cited
US3579495A Cited
US3763137A Cited
GB1532848A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1985-07-26 1986-01-25 1986-07-27 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1989-07-26 1990-01-25 1990-07-27 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1993-07-26 1994-01-25 1994-07-27 $7700.00 Window Closed / Lapsed

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