Application of reduced dyes in imaging

Patent metadata

Field	Value
Publication number	US-10100198-B2
Application number	US-201615258283-A
Country	US
Kind code	B2
Filing date	Sep 7, 2016
Priority date	Sep 23, 2011
Publication date	Oct 16, 2018
Grant date	Oct 16, 2018

Abstract

The present invention provides novel compounds and methods for hydrocyanines derived from near-infrared cyanine dyes, as reactive oxygen species probes in imaging. In certain embodiments, the present invention provides reduced dyes as substrates for ELISA and Western blots.

First claim

What is claimed is:

1. A compound, wherein the compound has the formula:

wherein R 1′ and R 1a′ are each independently hydrogen or deuterium;
M is an alkali metal cation; and
R 2 is CO 2 H or CO 2 M.

2. An assay method for measuring an analyte in a sample, the method comprising:
a) contacting said sample with a binding reagent that binds to an analyte to form an analyte complex;
b) contacting said sample with a compound of claim 1 , wherein said compound reacts with a radical, wherein said radical is generated from said analyte complex to form a detectable agent, which said detectable agent is a cyanine dye; and
c) upon excitation, detecting said detectable agent.

3. The method of claim 2 , wherein said analyte is a biological molecule.

4. The method of claim 2 , wherein said binding reagent is an antibody.

5. The method of claim 2 , wherein said analyte complex comprises an enzyme wherein a substrate of the enzyme is hydrogen peroxide.

6. The method of claim 2 , wherein said detectable agent is an oxidized form of a compound of claim 1 .

7. The method of claim 2 , wherein said assay method is an immunological detection method.

8. The method of claim 7 , wherein said immunological detection method is a member selected from the group consisting of an enzyme linked immunosorbent assay (ELISA), a Western blot analysis, and an immunohistochemical analysis.

9. A method for detecting a reactive oxygen species in a cell of a sample or a subject, said method comprising:
contacting a cell with one or more compounds of claim 1 , to form an oxidized form of said compound; and
exciting the oxidized form of said compound to emit light.

10. The method of claim 9 , wherein the oxidized form of said compound is detected by a method selected from the group consisting fluorescence spectroscopy or fluorescence microscopy.

11. A kit for conducting a fluorescent assay, said kit comprising a composition, said composition comprising:
a) a compound of claim 1 ; and
b) a buffer.

12. The kit of claim 11 , further comprising instructions for use of the composition in the fluorescent assay.

Assignees

• Li Cor Inc

Inventors

• Kundu Kousik
• Padhye Nisha
• Volcheck William

Classifications

• G01N33/575

  for cancer · CPC title

• C09B23/107

  four >CH- groups · CPC title

• C09B23/164 Primary

  containing one nitrogen atom · CPC title

• C07D209/08 Primary

  with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring · CPC title

• C09B23/086

  more than five >CH- groups · CPC title