Chemiluminescent methods and reagents for analyte detection

Abstract

The present invention relates to chemiluminescent method and regent to detect analyte. One aspect of the current invention relates to using enzyme substrate that can be cleaved by target enzyme to release chemiluminescent compound giving light signal for the detection of varieties of target enzymes. Another aspect of the current invention relates to use chemiluminescent enzyme coupled with analyte binding molecules to detect specific analyte molecules in a homogenous phase.

What is claimed is: 
     
         1 . A compound of formula I 
       
         
           
           
               
               
           
         
       
       where in T is a substituted or unsubstituted polycycloalkyl group bonded to the 4-membered ring portion of the dioxetane by a Spiro linkage; X is an aryl or heteroaryl moiety of 6-30 carbon atoms which induces chemiluminescent decomposition of the 1,2-dioxetane upon enzymatic cleavage of moiety Rs; R is an alkyl, aryl, aralkyl or cycloalkyl of 1-20 carbon atoms; Rs is selected from alpha-L-arabinopyranoside,beta-D-cellobioside,alpha-L-fucopyranoside, beta-D-fucopyranoside,beta-L-fucopyranoside,N-acetyl-alpha-D-galactosaminide, N-acetyl-beta-D-galactosaminide,alpha-D-galactopyranoside,beta-D-galactopyranoside, N-acetyl-alpha-D-glucosaminide,N-acetyl-beta-D-glucosaminide,alpha-D-glucopyranoside, beta-D-glucopyranoside,beta-D-glucuronic acid,beta-D-lactopyranoside,beta-D-maltopyranoside, alpha-D-mannopyranoside,beta-D-mannopyranoside and beta-D-xylopyranoside. 
     
     
         2 . A method to detecting analyte in a sample, comprising:
 a) contacting said sample with a first ligand of the analyte coupled with ATP producing enzyme and a second ligand of the analyte coupled with luciferase, wherein said luciferase use ATP as substrate; and   b) detecting the light generated from said luciferase.   
     
     
         3 . A kit for detecting analyte in a sample, comprising a first ligand of the analyte coupled with ATP producing enzyme and a second ligand of the analyte coupled with luciferase.