Detection and measurement of destructive and polymer forming enz

Details Detection and measurement of destructive and polymer forming enz Detection and measurement of destructive and polymer forming enz

1992-04-01

1994-05-10

Wityshyn, Michael G.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

435 23, 435962, 436811, C12Q 100, C12Q 137

Patent

active

053106474

ABSTRACT:
The change between a dispersed state and a flocculated state of a colloidal agent (e.g. Congo Rubin, colloidal gold) is used to provide sensitive visual detection and optionally assay of enzyme in a sample. The test cell includes a substrate for the enzyme, and, depending on the action of the enzyme, polymer which protects the colloid from electrolyte-induced flocculation is either formed or destroyed by the test reaction. In a test for a hydrolytic enzyme (e.g. a protease) the colloid loses protection and flocculates. In a test for a polymer-forming enzyme, the colloid gains protection and is prevented from flocculating by added electrolyte, or alternatively, depending on the polymer's behavior, becomes more sensitive to the flocculating action of the added electrolyte. The measurement may be quantitated using an instrumental monitor. The test uses a natural substrate for the enzyme to be assayed (e.g., gelatin for the gelatinase class of enzymes) and may achieve rapid speed. The test may be performed using a biologically-derived sample for diagnostically relevant purposes, e.g. to detect an enzyme indicative of periodontal disease. A kit for performing the test is also disclosed.

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