Use of signal enhancing compounds in electrochemiluminescence detection

Patent metadata

Field	Value
Publication number	US-9341575-B2
Application number	US-201313844778-A
Country	US
Kind code	B2
Filing date	Mar 15, 2013
Priority date	Oct 25, 2010
Publication date	May 17, 2016
Grant date	May 17, 2016

Abstract

The application provides methods for the detection of an analyte in a sample by electrochemiluminescence using certain reagent compositions. Reagent compositions, reagent kits for measuring electrochemiluminescence (ECL) and electrochemiluminescence detection methods using the reagent compositions are disclosed. In particular, the application relates to the use of novel combinations of compounds, which can be used in said measurements to provide improved ECL assay performance.

First claim

What is claimed is:

1. A method for detecting an analyte in a sample, comprising the steps of:
a) incubating the sample with a detection reagent labeled with an electrochemiluminescent group to provide an analyte-bound labeled detection reagent;
b) electrochemically triggering the release of electrochemiluminescence by the analyte-bound labeled detection reagent, which is incubated with a reagent composition for determining electrochemiluminescence (ECL), comprising:
i) at least one electrochemiluminescence (ECL) coreactant, which is selected from the group consisting of tertiary amines, oxalate and persulfate;
ii) a concentration of 0.01 M to 0.25 M of at least one carbonic acid amide selected from 2-pyrrolidone and compounds having a formula:

wherein R 1 is H, or a C 1 -C 5 alkyl group, optionally substituted with a single chlorine or fluorine atom, R 2 ═H, and R 3 ═H; and

c) determining the electrochemiluminescence (ECL) signal thereby detecting the analyte.

2. The method according to claim 1 , wherein the measurement of the analyte in the sample using ECL is performed in an aqueous solution.

3. The method according to claim 1 , wherein the at least one carbonic acid amide comprises acetamide, 2-fluoroacetamide, 2-chloroacteamide, propanamide, 2-chloropropanamide, 3-chloropropanamide, butanamide, 2-chlorobutanamide or a mixture thereof.

4. The method according to claim 1 , wherein the reagent composition further comprises a preservative.

5. The method according to claim 4 , wherein the reagent composition comprises 0.1% to 5% of the preservative.

6. The method according to claim 4 , wherein the preservative comprises boric acid, borate or a mixture thereof.

7. The method according to claim 1 , wherein the at least one ECL coreactant comprises a compound selected from the group consisting of tertiary amines, oxalate and persulfate.

8. A method for detecting an analyte in a sample, comprising the steps of:
a) incubating the sample with a detection reagent labeled with an electrochemiluminescent group to provide an analyte-bound labeled detection reagent,
b) electrochemically triggering the release of electrochemiluminescence by the analyte-bound labeled detection reagent, which is incubated with a reagent composition comprising:
i) at least one electrochemiluminescence (ECL) coreactant, which is selected from the group consisting of tertiary amines, oxalate and persulfate,
ii) a concentration of 0.01 M to 0.25 M of at least one carbonic acid amide selected from 2-pyrrolidone and compounds having a formula

wherein R 1 is —H, or a C 1 -C 5 alkyl group, optionally substituted with a single chlorine or fluorine atom,
R 2 ═H, and R 3 ═H; and

c) determining the electrochemiluminescence (ECL) signal thereby detecting the analyte.

9. The method of claim 8 , wherein R 1 is a C 1 -C 5 alkyl group.

10. The method of claim 8 , wherein R 1 is a monochloro-substituted C 1 -C 5 alkyl group.

11. The method of claim 8 , wherein the at least one carbonic acid amide comprises acetamide, 2-fluoroacetamide, 2-chloroacteamide, propanamide, 2-chloropropanamide, 3-chloropropanamide, butanamide, 2-chlorobutanamide or a mixture thereof.

12. The method of claim 8 , wherein the reagent composition further comprises boric acid, borate or a mixture thereof.

13. The method of claim 12 , wherein the reagent composition further comprises a detergent selected from the group consisting of fatty acid alcohol ethoxylates, alkylpolyglucosides, octylglucosides and mixtures thereof.

14. The method of claim 8 , wherein the reagent composition further comprises (a) boric acid, borate or a mixture thereof, (b) polidocanol and (c) or 4,4-dimethyl oxazolidine; and the ECL coreactant comprises tripropylamine.

15. The method of claim 8 , wherein the reagent composition comprises the at least one carbonic acid amide in a concentration of 0.01 M to 0.25 M;
the at least one carbonic acid amide comprises acetamide, 2-chloroacetamide, propanamide and butanamide; the ECL coreactant comprises tripropylamine; and
the reagent composition further comprises 0.1 to 5% boric acid, borate or a mixture thereof.

16. The method of claim 8 , wherein the labeled detection reagent is immobilized on a solid phase.

Assignees

• Roche Diagnostics Operations

Inventors

• Finke Andreas
• Hauptmann Bernhard
• Stoeckel Johannes
• Windfuhr Michaela

Classifications

• G01N21/66 Primary

  electrically excited, e.g. electroluminescence · CPC title

• B01L3/502761

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• B01L3/502715

  characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces · CPC title

• G01N21/76 Primary

  Chemiluminescence; Bioluminescence · CPC title

• C07D499/10 Primary

  Modification of an amino radical directly attached in position 6 · CPC title