Assay methods and systems

Abstract

Methods, systems, kits for carrying out a wide variety of different assays that comprise providing a first reagent mixture which comprises a first reagent having a fluorescent label. A second reagent is introduced into the first reagent mixture to produce a second reagent mixture, where the second reagent reacts with the first reagent to produce a fluorescently labeled product having a substantially different charge than the first reagent. A polyion is introduced into at least one of the first and second reagent mixtures, and the fluorescent polarization in the second reagent mixture relative to the first reagent mixture is determined, this fluorescent polarization being indicative of the rate or extent of the reaction.

Description

CROSS REFERENCE TO RELATED APPLICATION [0001] This application is a division of U.S. patent application Ser. No. 10,397,887, filed Mar. 26, 2003, which is a continuation of U.S. patent application Ser. No. 09 / 865,044, filed May 24, 2001, now U.S. Pat. No. 6,699,655, which is a continuation of U.S. patent application Ser. No. 09 / 316,447, filed May 21, 1999, now U.S. Pat. No. 6,287,774, each of which are incorporated herein in its entirety for all purposes.BACKGROUND OF THE INVENTION [0002] Virtually all chemical, biological and biochemical research depends upon the ability of the investigator to determine the direction of her research by assaying reaction mixtures for the presence or absence of a particular chemical species within the reaction mixture. In a simple case, the rate or efficiency of a reaction is assayed by measuring the rate of production of the reaction product, or the depletion of a reaction substrate. Similarly, interactive reactions, e.g., binding or dissociation re...

AI Technical Summary

Benefits of technology

The present invention provides methods, systems, kits, and assay systems for carrying out various assays using fluorescent labels. These assays include detecting reactions, identifying the presence of specific nucleotides, and detecting phosphorylation. The methods involve mixing reagents and a polyion, and measuring the fluorescence polarization to determine the reaction. The systems include a detection zone and a detector for measuring the fluorescence polarization. The kits include a volume of reagents and instructions for carrying out the assay. The invention allows for the efficient and accurate detection of reactions and provides a reliable method for measuring the reaction parameters.

Problems solved by technology

Unfortunately, many reactions of particular interest do not have the benefit of having a readily available surrogate reagent that produces signal only when subjected to the reaction of interest.

However, even when one member of the binding pair is labeled, the formation of the complex does not generally give rise to an optically detectable difference between the complex and the labeled molecule.

Similarly, there are a number of other assays whose reagents and / or products cannot be readily distinguished from each other, even despite the incorporation of optically detectable elements.

As should be apparent, assays requiring additional separation steps can be extremely time consuming and less efficient, as a result of losses during the various assay steps.

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