Method for enhancing enzyme assays

Abstract

Additives capable of controlling the reaction rate between substrate and enzyme to form a dye in enzyme assays with absorbance, chemiluminescence, and fluorescence are effective tools for optimizing enzyme assays. A dye can be formed rapidly from the reaction between enzyme and substrate in the presence of a catalyst. Using relatively high concentration of dye formed from the rapid reaction, trace levels of analytical materials can be quantified using absorbance, chemiluminecence and fluorescence detection. A dye can be formed from a relatively slow reaction between enzyme and substrate in the presence of a surfactant such as Triton® X-100 and β-cyclodextrin. Using relatively low concentration of dye from the slow reaction, a high concentration of analytical material can be quantified without any dilution using absorbance, chemiluminescence, and fluorescence.

What is claimed is: 
     
         1 . A method of enhancing an enzyme immunoassay, comprising reacting a substrate with an enzyme in the presence of a surfactant. 
     
     
         2 . The method of  claim 1 , wherein the enzyme immunoassay is ELISA, fluorescene EIA, Luminol CLEIA, 1,2-dioxetane CLEIA, ODB CLEIA, or ODI CLEIA. 
     
     
         3 . The method of  claim 1 , wherein the enzyme is horseradish peroxidase (HRP), Alkaline phophatase (ALP), glucose oxidase (GOx), β-Galactoxidase, or acetylcholinesterase (AchE). 
     
     
         4 . The method of  claim 1 , wherein the surfactant is anionic surfactant, cationic surfactant, or non-ionic surfactant. 
     
     
         5 . The method of  claim 4 , wherein the anionic surfactant is Sodium dodecyl sulfate. 
     
     
         6 . The method of  claim 4 , wherein the cationic surfactant is Hexadecyltrimethylammonium bromide, or Hexadecyltrimethylammonium chloride. 
     
     
         7 . The method of  claim 4 , wherein the non-ionic surfactant is a Triton X derivative or a cyclodextrin derivative. 
     
     
         8 . The method of  claim 7 , wherein the Triton X derivative is 4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol, 4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol, (1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol, Polyethylene glycol tert-octylphenyl ether, or Octylphenol Ethoxylate. 
     
     
         9 . The method of  claim 7 , wherein the cyclodextrin derivative is α-cyclodextrin, (2-Hydroxypropyl)-β-cyclodetrin, β-cyclodextrin, Carboxymethyl-β-cyclodextrin, Carbomethyl-β-cyclodextrin sodium salt, or γ-cyclodextrin.