Reagents and methods for the rapid and quantitative assay of pharmacological agents
First Claim
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1. An assay for determining the presence of a target analyte in a test sample comprising the steps of:
- (A) forming a reaction mixture by contacting a test sample with;
(i) a bidentate reagent comprising a biotin member, an analyte member, and a spacer member between said biotin and analyte members, wherein;
(a) said biotin member of said reagent is bound to a biotin-binding agent selected from the group consisting of avidin and streptavidin;
said biotin-binding agent being covalently bound to a solid support, said solid support being a latex particle having a particle size of about 38 to 100 nanometers;
(b) said analyte member of said reagent is capable of specifically binding to an antibody capable of binding to a target analyte;
(c) said intermediate spacer member is sufficient in length to permit said analyte member to bind to the antibody and said biotin member to bind to the biotin-binding agent; and
(ii) said antibody;
(B) incubating said reaction mixture under conditions sufficient to permit the formation of a complex between said bidentate reagent and said antibody; and
(C) measuring the extent of any formation of said complex using nephelometric or turbidimetric means, said extent being inversely proportional to the concentration of said target analyte in said sample.
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Abstract
Bidentate reagents for rapidly and quantitatively assaying the concentration of pharmacological agents in biological samples are described. The reagents are used in an immunoassay format for determining the concentration of desired, preselected pharmacological agents, such as benzoylecgonine, cocaine, an opiate, PCP, digoxigenin, acetaminophen, carbamazepine, phenytoin, primidone, theophylline, an aminoglycoside antibiotic, vancomycin, quinidine or a cannabinoid.
52 Citations
25 Claims
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1. An assay for determining the presence of a target analyte in a test sample comprising the steps of:
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(A) forming a reaction mixture by contacting a test sample with; (i) a bidentate reagent comprising a biotin member, an analyte member, and a spacer member between said biotin and analyte members, wherein; (a) said biotin member of said reagent is bound to a biotin-binding agent selected from the group consisting of avidin and streptavidin;
said biotin-binding agent being covalently bound to a solid support, said solid support being a latex particle having a particle size of about 38 to 100 nanometers;(b) said analyte member of said reagent is capable of specifically binding to an antibody capable of binding to a target analyte; (c) said intermediate spacer member is sufficient in length to permit said analyte member to bind to the antibody and said biotin member to bind to the biotin-binding agent; and (ii) said antibody; (B) incubating said reaction mixture under conditions sufficient to permit the formation of a complex between said bidentate reagent and said antibody; and (C) measuring the extent of any formation of said complex using nephelometric or turbidimetric means, said extent being inversely proportional to the concentration of said target analyte in said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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Specification