Dna fragmentation assay
AssigneeSanofi-Aventis
InventorsTina Garyantes, Zhuyin Li, Yongping Yan, Justin Anthony Yu, Asher Zilberstein
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Abstract
The present invention provides methods for the detection of agents that modify the formation of DNA fragmentation in cells. The disclosed methods are configured in an assay format amendable to high throughput screening applications.
Claims26
A method of identifying an agent that modifies the formation of DNA fragments, the method comprising:
- a) providing cells in an array of receptacles;
- b) adding an agent to at least one receptacle;
- c) incubating the agent with the cells for a predetermined period of time;
- d) lysing the cells;
- e) adding a detectable compound capable of intercalating into DNA fragments to said at least one receptacle;
- f) measuring the amount of detectable compound intercalated;
- and
- g) comparing the amount of intercalated detectable compound to a control to determine a difference
- thereby identifying said agent as a modifying agent when the difference exceeds a predetermined threshold.
A method of identifying an agent that modifies the formation of DNA fragments, the method comprising:
- a) providing cells in an array of receptacles;
- b) adding to at least one receptacle a component selected from the group consisting of an inducer, an inhibitor, a modulator, a modulator of the inducer and a modulator of the inhibitor;
- c) incubating the component with the cells for a predetermined period of time;
- d) adding an agent to said at least one receptacle;
- e) incubating the agent with the cells for a predetermined period of time;
- f) lysing the cells;
- g) adding a detectable compound capable of intercalating into DNA fragments to said at least one receptacle;
- h) measuring the amount of detectable compound intercalated;
- and
- i) comparing the amount of intercalated detectable compound to a control to determine a difference
- thereby identifying said agent as a modifying agent when the difference exceeds a predetermined threshold.
A method of identifying an agent that modifies the formation of DNA fragments, the method comprising
- a) providing cells in an array of receptacles;
- b) adding an agent to at least one receptacle;
- c) incubating the agent with the cells for a predetermined period of time;
- d) adding a detectable compound capable of intercalating into DNA fragments to said at least one receptacle;
- e) measuring the amount of detectable compound intercalated;
- and
- f) comparing the amount of intercalated detectable compound to a control to determine a difference
- thereby identifying said agent as a modifying agent when the difference exceeds a predetermined threshold.
A method of identifying an agent that modifies the formation of DNA fragments, the method comprising:
- a) providing cells in an array of receptacles;
- b) adding to at least one receptacle a component selected from the group consisting of an inducer, an inhibitor, a modulator, a modulator of the inducer and a modulator of the inhibitor;
- c) incubating the component with the cells for a predetermined period of time;
- d) adding an agent to said at least one receptacle;
- e) incubating the agent with the cells for a predetermined period of time;
- f) adding a detectable compound capable of intercalating into DNA fragments to said at least one receptacle;
- g) measuring the amount of detectable compound intercalated;
- and
- h) comparing the amount of intercalated detectable compound to a control to determine a difference
- thereby identifying said agent as a modifying agent when the difference exceeds a predetermined threshold.
An assay system for identifying an agent that modifies the formation of DNA fragments, the assay system comprising:
- a) an array of receptacles;
- b) a lysis buffer;
- c) a detectable compound capable of intercalating into DNA;
- and
- d) at least one component wherein the component is selected from the group consisting of the agent(s), an inducer(s), an inhibitor, a modulator(s), a modulator(s) of the inducer(s), a modulator(s) of the inhibitor(s), control(s) and cells.
A method of diagnosing or monitoring a treatment of a disease wherein a biomarker for the disease comprises the formation of DNA fragments, the method comprising:
- a) providing a biological sample in an array of receptacles;
- b) adding a detectable compound capable of intercalating into DNA fragments to at least one receptacle;
- c) measuring the amount of detectable compound intercalated;
- and
- d) comparing the amount of intercalated detectable compound to a reference to determine a difference
- thereby diagnosing or monitoring the treatment of the disease when the difference exceeds a predetermined threshold.
An assay system for diagnosing or monitoring a treatment of a disease wherein a biomarker for the disease comprises the formation of DNA fragments, the assay system comprising:
- a) an array of receptacles;
- b) a detectable compound capable of intercalating into DNA;
- and
- d) at least one control.
Dependencies are inferred from the claim text.