Compositions and methods for simultaneous inactivation of alkaline phosphatase and peroxidase enzymes during automated multiplex tissue staining assays
AssigneeVentana Medical Systems, Inc.
InventorsJames Grille, Brian D. Kelly, Jerome Kosmeder, II, Eric May, Noemi Sebastiao, Pamela Wirth
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Abstract
Disclosed are compositions and methods for inactivating one or more enzymes in a biological sample.
Claims20
A method of detecting targets in a biological sample, comprising:
- a) contacting the biological sample with one or more detection reagents, each detection reagent comprising a different enzyme;
- b) detecting signals from the one or more detection reagents;
- and
- c) inactivating each of the different enzymes of the one or more detection reagents by dispensing an enzyme inactivation composition to the biological sample, wherein the first enzyme inactivation composition comprises a polycarboxylic acid having a pH ranging from about 1 to about 3;
- and optionally at least one of a peroxide or a preservative, wherein at least one of the first enzyme inactivation composition or the biological sample is maintained at a first temperature ranging from between about 25° C. to about 50° C. for a first time period ranging from between about 4 minutes to about 16 minutes.
A biological sample comprising one or more enzymes that are either substantially inactivated or completely inactivated, the biological sample prepared by introducing to the biological sample an enzyme inactivation composition comprising a polycarboxylic acid and hydrogen peroxide, wherein the hydrogen peroxide is present in an amount of about 0.25% to about 5% by total weight of the enzyme inactivation composition, and wherein the enzyme inactivation composition is allowed to remain in contact with the biological sample for at least about 4 minutes at a temperature of between about 25° C. and about 50° C.
An enzyme inactivation composition comprising a polycarboxylic acid having a pH ranging from about 1 to about 3;
- a peroxide;
- and a preservative, wherein the peroxide is present in an amount ranging from between about 0.25% to about 5% by total weight of the composition, and wherein the preservative is present in an amount ranging from between about 0.05% to about 1.0% by total weight of the composition.
Dependencies are inferred from the claim text.