US3616231A
Expired / Lapsed
US
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Process for the production of uricase
Assignee: Roche Diagnostics GmbH
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Filing: 1968-11-14
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Granted: 1971-10-26
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Expires: 1988-11-14
Abstract
Abstract Process for isolating uricase from animal tissue such as liver, kidney and spleen comprising extracting the water-insoluble fraction of such a tissue homogenate with an alkaline salt solution, treating the extract containing the uricase with kieselguhr (diatomite), separating the kieselguhr having the uricase adsorbed thereon from the remaining solution, eluting the separated kieselguhr with an alkaline acetone solution, precipitating the uricase in the eluate with ammonium sulfate and purifying the recovered precipitate by washing the same at least once with water or water containing a minor amount of a salt.
Inventors
- 👤 Hans Ulrich Bergmeyer
- 👤 Hans Mollering
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 16:49
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0012; C12N9/0044; C12N9/0046; Y; Y10; Y10S; Y10S435/00; Y10S435/814; Y10S435/815
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
23 words
Process according to claim 1 wherein said tissue comprises at least one member of the group consisting of animal liver, kidney and spleen.
Claim 2
Dependent Claim
41 words
Process according to claim 1 which comprises effecting said contacting of the alkaline salt solution containing uricase with the kieselguhr of step (b) at a pH of between 6 and 8 and a salt concentration of between 0.1 and 0.5 M.
Claim 3
Dependent Claim
40 words
Process according to claim 1 which comprises effecting said contacting of the alkaline salt solution containing uricase with the kieselguhr of step (b) at a pH of 6.7 to 7.3 and a salt concentration of between 0.1 and 0.5 M.
Claim 4
Dependent Claim
43 words
Process according to claim 1 which comprises effecting said extracting of the kieselguhr having uricase adsorbed thereon with alkaline acetone solution of step (c) with an aqueous acetone solution containing 20 to 35 percent acetone and having a pH of 8.0 to 9.8.
Claim 5
Dependent Claim
56 words
Process according to claim 1 which comprises adjusting the alkaline acetone extract obtained in step (c) to an acetone concentration of about 60 percent to thereby precipitate the uricase from said extract, dissolving the uricase precipitate with an alkaline salt solution and thereafter treating the resulting solution with ammonium sulfate to precipitate the uricase present therein.
Claim 6
Dependent Claim
29 words
Process according to claim 6 wherein the dissolution of the uricase precipitate is carried out with a dilute alkali carbonate solution having a pH of from 9 to 10.5.
Claim 7
Dependent Claim
26 words
Process according to claim 6 wherein the dissolution of the uricase precipitate is carried out with a dilute alkali carbonate solution having a pH of 10.2.
Claim 8
Dependent Claim
24 words
Process according to claim 1 which comprises extracting the ammonium sulfate precipitate in step (e) with an 0.01 to 0.3 M potassium carbonate solution.
Claim 9
Dependent Claim
22 words
Process according to claim 1 which comprises extracting the ammonium sulfate precipitate in step (e) with an 0.05 M potassium carbonate solution.
Claim 10
Dependent Claim
69 words
Process according to claim 1 wherein the extraction of step (e) is carried out at least two times and wherein following said first extraction, the uricase precipitate is dissolved in an aqueous alkaline solution and the solution treated with solid calcium phosphate gel to remove any colored impurities, the uricase precipitated out of solution with ammonium sulfate and the precipitate subjected to a second extraction according to step (e).
Claim 11
Dependent Claim
24 words
Process according to claim 11 wherein said uricase precipitate following said first extraction is dissolved in 0.2 M Na2CO3 having a pH of 10.2.
Claim 12
Dependent Claim
23 words
Process accordinG to claim 1 which comprises effecting the ammonium sulfate precipitation of step (d) with an ammonium salt concentration of 1.6 M.
Claim 13
Dependent Claim
29 words
Process according to claim 1 wherein said water-insoluble fraction of animal tissue homogenate is obtained by extracting homogenized animal tissue with aqueous glycine-Na2CO3 buffered to a pH of 10.2.
Claim 14
Dependent Claim
12 words
Process according to claim 1 wherein said animal tissue is pig liver.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1974-10-25 | 1975-04-26 | 1975-10-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1978-10-25 | 1979-04-26 | 1979-10-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1982-10-25 | 1983-04-26 | 1983-10-26 | $7700.00 | Window Closed / Lapsed |