Assays for measuring nucleic acid damaging activities
AssigneeIGEN International, Inc.
InventorsJeffrey A. Heroux, George B. Sigal, Reid W. von Borstel
Life of this patent
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Abstract
A method for assaying a sample for a nucleic acid damaging activity using at least one singular double-stranded nucleic acid with at least one electrochemiluminescent label, and a method for measuring an inhibitor of a nucleic acid damaging activity with at least one singular double-stranded nucleic acid using at least one electrochemiluminescent label, are disclosed.
Claims26
A method for assaying a sample for the presence of a nucleic acid repair protein, which comprises:
- a) mixing at least one predetermined single- or double-stranded nucleic acid containing at least one label and containing a nucleic acid repair protein binding domain with a sample which may contain a nucleic acid repair protein,
- b) incubating the mixture of step a) under conditions which allow the binding of said nucleic acid repair protein to said at least one predetermined single- or double-stranded nucleic acid repair protein binding domain to form a complex,
- c) adding a nucleic acid-cleaving enzyme or reagent to the mixture of step b),
- d) incubating the mixture of step c) under conditions which allow the cleavage of said at least one predetermined single- or double-stranded nucleic acid which has not formed a complex, and
- e) measuring the amount of said complex to measure said nucleic acid repair protein.
A method for assaying a sample for the presence an inhibitor of a nucleic acid repair protein, which comprises:
- a) mixing at least one predetermined single- or double-stranded nucleic acid containing at least one label and containing a nucleic acid repair protein binding domain and a predetermined nucleic acid repair protein with a sample which may contain an inhibitor of the binding of said nucleic acid repair protein with said at least one predetermined single- or double-stranded nucleic acid,
- b) incubating the mixture of step a) under conditions which allow the binding of said nucleic acid repair protein to said at least one predetermined single- or double-stranded nucleic acid to form a complex,
- c) adding a nucleic acid-cleaving enzyme or reagent to the mixture of step b),
- d) incubating the mixture of step c) under conditions which allow the cleavage of said at least one predetermined single- or double-stranded nucleic acid which has not formed a complex, and
- e) measuring the amount of said complex to measure said inhibitor.
A method for assaying a sample for a nucleic acid damaging activity which comprises:
- a) mixing at least one single- or double-stranded nucleic acid containing at least one electrochemiluminescent label with a sample which may contain a nucleic acid-damaging activity,
- b) incubating the mixture of step (a) under conditions which allow said nucleic acid damaging activity to damage said nucleic acid,
- c) adding a reagent that specifically cleaves, damaged nucleic acid,
- d) incubating the mixture of step (c) under conditions that allow the cleavage of damaged nucleic acid, and
- e) measuring the amount of cleaved nucleic acid to measure said nucleic acid damaging activity.
A method for measuring an inhibitor of a nucleic acid damaging activity, which comprises:
- a) mixing at least one single- or double-stranded nucleic acid containing at least one electrochemiluminescent label with a predetermined nucleic acid damaging activity, and a sample which may contain an inhibitor of said nucleic acid damaging activity,
- b) incubating the mixture of step (a) under conditions which allow said nucleic acid damaging activity to damage said nucleic acid,
- c) adding a reagent that specifically cleaves damaged nucleic acid,
- d) incubating the mixture of step (c) under conditions that allow the cleavage of damaged nucleic acid, and
- e) measuring the amount of cleaved nucleic acid to measure said inhibitor of said nucleic acid damaging activity.
A kit comprising in one or more containers:
- a) a nucleic acid having a predetermined nucleic acid repair protein binding domain wherein said nucleic acid has a detectable moiety attached thereto, and
- b) a nucleic acid-cleaving enzyme or nucleic acid-cleaving reagent.
A nucleic acid comprising a predetermined nucleic acid repair protein binding domain wherein said nucleic acid has a detectable moiety attached thereto.
Dependencies are inferred from the claim text.