Ligand analog-irreversible enzyme inhibitor conjugates and metho

Details Ligand analog-irreversible enzyme inhibitor conjugates and metho Ligand analog-irreversible enzyme inhibitor conjugates and metho

1979-02-05

1981-06-16

Wiseman, Thomas G.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving antigen-antibody binding, specific binding protein...

435 20, 435184, 435810, 424 12, 23230B, G01N 3354, C12N 999

Patent

active

042738660

ABSTRACT:
The present invention encompasses a method for determining ligands in test samples comprising intermixing with the test sample a ligand analog-irreversible enzyme inhibitor conjugate and a binding protein bindable to the ligand and the ligand analog-irreversible enzyme inhibitor conjugate and wherein the amount of ligand analog-irreversible enzyme inhibitor conjugate bound by the binding protein is related to the amount of ligand in the test sample, said binding protein inactivating the irreversible enzyme inhibitor when bound to the ligand analog portion of the conjugate; intermixing an enzyme which is irreversibly inhibited by the ligand analog-irreversible enzyme inhibitor conjugate unbound by the binding protein; and intermixing substrate to the enzyme and monitoring the enzyme substrate reaction.
The invention also includes ligand analog-irreversible enzyme inhibitor conjugates useful as reagents in practicing the method. Methods and reagents of the present are particularly useful in determining drugs, hormones, and the like in biological fluids.

REFERENCES:
patent: 4120754 (1978-10-01), Barendz et al.
patent: 4134792 (1979-01-01), Boguslaski et al.
Aaron et al., "The Stereochemistry of Asymmetric Phosphorus Compounds II. Stereospecificity in the Irreversible Inactivation of Cholinesterases by the Enantiomorphs of an Organophosphorus Inhibitor," J. Am. Chem. Soc., vol. 80 (1958) pp. 456-458.
Aaron et al., "The Stereochemistry of Asymmetric Phosphorus Compounds. III. The Restoration of a Series of O-Alkylphosphonothioic Acids," J. Am. Chem. Soc., vol. 82 (1960) pp. 596-598.
Boter et al., "Organophosphorus Compounds. Part V. The Resolution of O-Alkyl Hydrogen Methylphosphonothioates with (+) and (-)-.alpha.-Phenylethylamine", Rec. Trav. Chim., vol. 86 (1967) pp. 399-404.
Aldridge et al., Enzyme Inhibitors as Substrates, American Elsevren Publishing Co., New York, (1972) pp. 5, 6.
Pilz, "Cholinesterases", Methods of Enzymatic Analysis, vol. 2, Academic Press, New York (1974) Bergmeyer Ed. pp. 831-835, 851-855.

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