Optical method and device for identifying and quantifying analytes

• US 9,846,154 B2
• Filed: 12/20/2012
• Issued: 12/19/2017
• Est. Priority Date: 12/20/2011
• Status: Active Grant

• Current Assignee: Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Original Assignee: Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Inventors: Surez, Guillaume, Santschi, Christian, Martin, Olivier
• Primary Examiner(s): Crawford, Erik B
• Application Number:
  US14/366,597
• Publication Number:
  US 20140322733A1
• Time in Patent Office: 1,825 Days
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: G01N 21/314 with comparison of measurements at specific and non-specific wavelengths dual wavelength spectrometry G01J3/427
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes

Abstract

A method for analyzing a metalloprotein and/or the interaction with its environment comprising the following steps: (a) Providing a medium that enhances the detection of the electromagnetic cross-section signal of metalloproteins, (b) Incorporating a metalloprotein to analyse into said medium, (c) Contacting said medium with electromagnetic radiation, (d) Obtaining the electromagnetic cross-section spectrum of said metalloprotein, (e) Determining from said electromagnetic cross-section spectrum at least one parameter related to one or several analytes of interest.

Claims

1. A method for analyzing an analyte comprising the steps of:

(a) providing a multiscattering medium that enhances a detection of an electromagnetic cross-section signal of a metalloprotein,

(b) incorporating the metalloprotein to be analyzed into the multiscattering medium and the analyte is arranged to establish a redox reaction with the metalloprotein,

(c) contacting the multiscattering medium with electromagnetic radiation,

(d) obtaining the electromagnetic cross-section spectrum of the metalloprotein, and

(e) determining from the electromagnetic cross-section spectrum a concentration of the analyte.

2. The method according to claim 1 wherein the electromagnetic cross-section is an absorption cross-section of the metalloprotein.

3. The method according to claim 1 wherein the multiscattering medium is at least one of an aggregate;

a suspension;

a gel;

a porous membrane;

a microstructured substrate; and

a nanostructured substrate.

4. The method according to claim 1 wherein the analyte is a reaction product of enzymatic or chemical reactions.

5. The method according to claim 1 wherein the step of (c) contacting uses a dark-field electromagnetic radiation illumination.

6. The method according to claim 1 wherein the step of (e) determining further includes:

determining the concentration of the analyte based on a measurement of a molecular state of the metalloprotein.

7. The method according to claim 6 wherein the molecular state is an oxidation state.

8. The method according to claim 1 wherein the analyte is hydrogen peroxide.

9. The method according to claim 1 wherein the metalloprotein is a cytochrome c.

10. The method according to claim 1, wherein the multiscattering medium enhances an optical cross-section of the metalloprotein.

11. The method according to claim 1, wherein the multiscattering medium provides for an amplification of the electromagnetic cross-section spectrum to suppress a background signal.