US4066509A
Expired / Lapsed
US
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Detection of hydrolyzing enzymes
Assignee: Roche Diagnostics GmbH
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Filing: 1974-02-08
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Granted: 1978-01-03
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Expires: 1994-02-08
Abstract
Abstract The activity, or concentration, of a hydrolyzing enzyme, e.g., an endo-enzyme, is determined by providing a detection layer comprising a carrier matrix and a water-insoluble substrate dispersed in said carrier matrix which substrate is detectably marked, e.g., dyed, and which substrate, upon action of a hydrolyzing enzyme thereon, is broken down into soluble fragments capable of diffusing through the carrier matrix to cause formation of an unmarked area in said detection layer; and measuring the size of the unmarked area as a measure of the activity of concentration of said enzyme.
Inventors
- 👤 Miroslav Ceska
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 19:13
CPC Classifications
G; G01; G01N; G01N31/00; G01N31/22; C; C12; C12Q; C12Q1/00; C12Q1/34; G01N2333/00; G01N2333/90; G01N2333/914; G01N2333/924; G01N2333/948; G01N2333/95; G01N2333/964; G01N2333/96425; G01N2333/96427; G01N2333/9643; G01N2333/96466; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/8215; Y10S435/822; Y10S435/832; Y10S435/839; Y10S435/911; Y10S435/933
Claims Analysis
Total Claims: 4
Claim 1
Independent Claim
128 words
Method for the detection of the activity, or concentration, of a hydrolyzing enzyme selected from the group consisting of alpha-amylase, dextranase, and papain, which method comprises applying a solution of said hydrolyzing enzyme onto a detection layer consisting essentially of a carrier matrix gel inert to the enzyme and permitting diffusion of substrate fragments therein and, dispersed throughout said matrix gel, a dyed intermolecularly cross-linked substrate selected from the group consisting of starch, dextran, cellulose and albumin, cross-linked with a diepoxide to render the substrate insoluble in the matrix gel, and measuring the size of an unmarked area formed in said gel due to fragmentation and diffusion of the dyed substrate by action of the enzyme thereon, as a measure of the amount of concentration of said enzyme.
Claim 2
Dependent Claim
128 words
Method for the detection of the activity, or concentration, of a hydrolyzing enzyme selected from the group consisting of alpha-amylase, dextranase, and papain, which method comprises applying a solution of said hydrolyzing enzyme onto a detection layer consisting essentially of a carrier matrix gel inert to the enzyme and permitting diffusion of substrate fragments therein and, dispersed throughout said matrix gel, a dyed intermolecularly cross-linked substrate selected from the group consisting of starch, dextran, cellulose and albumin, cross-linked with a diepoxide to render the substrate insoluble in the matrix gel, and measuring the size of an unmarked area formed in said gel due to fragmentation and diffusion of the dyed substrate by action of the enzyme thereon, as a measure of the amount of concentration of said enzyme.
Claim 3
Dependent Claim
17 words
Method as claimed in claim 1 wherein said matrix gel is an agar or a starch gel.
Claim 4
Dependent Claim
77 words
Method as claimed in claim 1 wherein the time required for producing the unmarked area after complete action of the enzyme on the substrate is first determined for a specific enzyme/detection layer combination under specific conditions and said hydrolyzing enzyme is then determined by measuring said unmarked area after a predetermined time shorter than the first determined time and comparing the unmarked area with a reference standard to calculate the amount, or concentration, of said hydrolyzing enzyme.
References Cited (Backward Citations)
US3416998A
Cited
US3676303A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-01-02 | 1981-07-04 | 1982-01-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-01-02 | 1985-07-04 | 1986-01-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-01-02 | 1989-07-04 | 1990-01-03 | $7700.00 | Window Closed / Lapsed |