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Last Updated: March 28, 2024

Claims for Patent: 4,804,625


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Summary for Patent: 4,804,625
Title: Assay procedures
Abstract:Binding assay methods involving determining the presence of analytes in samples through enzymatic formation of detectable substances in amounts related to the amount of analyte present in the sample and monitoring for the presence of the substances in distinct phases. Methods according to the invention include use of labelled materials which associate with the analyte to be determined or compete with the analyte for association with an added binder. The labelled materials employed include label portions which enzymatically form substances from substrates provided in or existing as a first phase, or, upon enzymatic treatment in a first phase, disassociate into substances capable of existing in or as a second distinct phase. Formation of the detectable substances is monitored by determining the transfer of the substance to a second distinct phase in contact with the first phase or by determining formation of a second distinct phase. The assays are useful in determining human IgG protein in blood samples and other constituents of blood or other biological samples without elaborate instrumentation, allowing for practice outside the clinical laboratory.
Inventor(s): Morrison; Larry E. (Lisle, IL), Royer; Garfield P. (Warrenville, IL), Heller; Michael J. (Poway, CA)
Assignee: Amoco Corporation (Chicago, IL)
Application Number:06/656,011
Patent Claims:1. A method for determining the presence of an analyte in a sample comprising contacting a sample with a reagent adapted to form a substrate phase and a product phase with said sample, said reagent including an enzyme-substrate pair, including an enzyme and a substrate, and labeled material, capable of assuming a bound position with said analyte, said labeled material having a label moiety capable of participating in reactions related to the presence of said analyte which reactions require said substrate present in said substrate phase and produce a reaction product, said label moiety is a member of said enzyme-substrate pair, said enzyme catalyzing a reaction with said substrate which produces said reaction product, said enzyme-substrate pair is selected from the group of enzyme-substrate pairs consisting of alkaline phosphatase and the substrate p-phenylazophenyl phosphate, and the enzyme collagenase and the substrate collagen/blue dextran, said reaction product capable of partitioning into said product phase distinct from said substrate phase wherein said substrate phase is a solid or a liquid, and said product phase is a liquid; and monitoring said product phase for the presence of said reaction product.

2. The method of claim 1 wherein said analyte and labeled material are members of a biological binding pair.

3. The method of claim 2 wherein said biological binding pair is selected from the group of biological binding pairs consisting of antigens and antibodies, complementary polynucleotides, drugs and drug receptors, and enzyme and enzyme substrates.

4. A kit for determining the presence of an analyte in a sample comprising: reagent adapted to form a substrate phase and a product phase with said sample, said reagent including an enzyme-substrate pair, including an enzyme and a substrate, and labeled material, capable of assuming a bound position with said analyte, said labeled material having a label moiety capable of participating in reactions related to the presence of said analyte which reactions require a substrate present in said substrate phase and produce a reaction product, said label moiety is a member of said enzyme-substrate pair, said enzyme catalyzing a reaction with said substrate which produces said reaction product, said enzyme substrate pair is selected from the group of enzyme-substrate pairs consisting of the enzyme alkaline phosphatase and the substrate p-phenylazophenyl phosphate, the enzyme collagenase and the substrate collagen/blue dextran, said reaction product capable of partitioning into said product phase distinct from said substrate phase wherein said substrate phase is a solid or a liquid and said product phase is a liquid; said reaction product detectable within said product phase to indicate the presence of said analyte.

5. The kit of claim 4 wherein said analyte and labeled material are members of a biological binding pair.

6. The kit of claim 5 wherein said biological binding pair is selected from the group of biological binding pairs consisting of antigens and antibodies, complementary polynucleotides, drugs and drug receptors, and enzyme and enzyme substrates.

7. A method for determining the presence of an analyte in a sample comprising contracting a sample with a reagent adapted to form a substrate phase and a product phase with said sample, said reagent including an enzyme-substrate pair, including an enzyme and a substrate, and labeled material, capable of assuming a bound position with said analyte, said labeled materials having a label moiety capable of participating in reactions related to the presence of said analyte which reactions require said substrate present in said substrate phase and produce a reaction product, said product capable of partitioning into said product phase distinct from said substrate phase wherein said substrate phase is a solid or a liquid, and said product phase is a gas and said product is detectable by olfactory means; and monitoring said product phase for the presence of said reaction product.

8. The method of claim 7 wherein said label moiety is a member of an enzyme-substrate pair, said enzyme catalyzing a reaction with said substrate which products said product.

9. The method of claim 8 wherein said enzyme substrate pair includes the enzyme peroxidase and the substrate 2-methyl-3-oxobutanal.

10. A kit for determining the presence of an analyte in a sample comprising: reagent including labeled material capable of assuming a bound position with said analyte, said labeled material having a label moiety capable of participating in reactions related to the presence of said analyte which produce a reaction product, and said label moiety is a member of an enzyme-substrate which includes the enzyme peroxidase and the substrate 2-methyl-3-oxobutanal, said reaction product being detectable to indicate the presence of said analyte.

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