Assays for measuring nucleic acid binding proteins and enzyme activities

Abstract

Processes for measuring DNA or RNA binding proteins, specific nucleic acids, as well as enzyme activities using labeled nucleic acids of labeled protein/peptide molecules are provided.

1 .- 2 . (canceled) 
     
     
         3 . A method of assaying a sample for an enzyme activity that cleaves nucleic acid, comprising:
 a) mixing at least one predetermined single- or double-stranded nucleic acid containing at least one electrochemiluminescent label with a sample, wherein the sample may contain a nucleic acid-cleaving enzyme;   b) incubating the mixture of step (a) under conditions which allow cleavage of the nucleic acid; and   c) analyzing the incubated mixture for cleaved nucleic acid.   
     
     
         4  The method of  claim 3 , wherein the electrochemiluminescent label comprises ruthenium, osmium or rhenium. 
     
     
         5 . The method of  claim 3 , wherein the electrochemiluminescent label comprises a bipyridyl or phenanthrolyl-containing complex of ruthenium. 
     
     
         6 . The method of  claim 3 , wherein the electrochemiluminescent label comprises RuBpy. 
     
     
         7 . The method of  claim 3 , wherein analyzing the incubated mixture comprises measuring the amount of cleaved nucleic acid in the incubated mixture. 
     
     
         8 . The method of  claim 3 , wherein the nucleic acid-cleaving enzyme is a nuclease. 
     
     
         9 . The method of  claim 8 , wherein the nuclease is a ribonuclease. 
     
     
         10 . The method of  claim 8 , wherein the nuclease is a deoxyribonuclease. 
     
     
         11 . The method of  claim 3 , wherein the nucleic acid-cleaving enzyme is an integrase. 
     
     
         12 . The method of  claim 3 , wherein the at least one electrochemiluminescent label is linked to the single- or double-stranded nucleic acid by a covalent bond or a specific interaction with a binding group. 
     
     
         13 . The method of  claim 3 , wherein analyzing the incubated mixture comprises analyzing an incubated mixture, which includes an ECL coreactant. 
     
     
         14 . The method of  claim 13 , wherein the ECL coreactant comprises a tertiary amine. 
     
     
         15 . The method of  claim 3 , wherein the nucleic acid has a capture moiety attached thereto. 
     
     
         16 . The method of  claim 15 , wherein the capture moiety is selected from the group consisting of antibody, antigen, receptor, ligand, and a nucleic acid base pairing polymer. 
     
     
         17 . A method of detecting an inhibitor of a nucleic acid-cleaving enzyme, comprising:
 a) combining to form a mixture i) at least one predetermined single- or double-stranded nucleic acid containing at least one electrochemiluminescent label, ii) at least one predetermined nucleic acid-cleaving enzyme, and iii) a sample, wherein the sample may contain an inhibitor of the nucleic acid-cleaving enzyme;   b) incubating the mixture of step (a) under conditions which allow cleavage of said nucleic acid; and   c) analyzing the incubated mixture for cleaved nucleic acid.   
     
     
         18 . The method of  claim 17 , wherein the electrochemiluminescent label is linked to the single- or double-stranded nucleic acid by a covalent bond or a specific interaction with a binding group. 
     
     
         19 . The method of  claim 17 , wherein the electrochemiluminescent label comprises a bipyridyl or phenanthrolyl-containing complex of ruthenium. 
     
     
         20 . The method of  claim 17 , wherein analyzing the incubated mixture comprises measuring the amount of cleaved nucleic acid in the incubated mixture. 
     
     
         21 . The method of  claim 17 , wherein the nucleic acid-cleaving enzyme is a nuclease. 
     
     
         22 . The method of  claim 17 , wherein the nucleic acid-cleaving enzyme is an integrase.