Method for assaying a target analyte while reducing interference in an immunosensor system

• US 7,910,352 B2
• Filed: 02/18/2010
• Issued: 03/22/2011
• Est. Priority Date: 09/10/2003
• Status: Active Grant

• Current Assignee: Abbott Point Of Care Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: Abbott Point Of Care Incorporated (Abbott Laboratories Incorporated)
• Inventors: Miller, Cary James, Campbell, John Lewis Emerson
• Primary Examiner(s): BROWN, MELANIE YU
• Application Number:
  US12/708,429
• Publication Number:
  US 20100167312A1
• Time in Patent Office: 397 Days
• Field of Search: 422/50, 422/55, 422/57, 422/58, 422/61, 422/68.1, 422/82.05, 422/82.01, 435/7.1, 435/7.7, 435/283.1, 435/287.1, 435/288.7, 436/518, 436/524, 436/525, 436/528
• US Class Current: 435/287.1
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

An electrochemical immunosensor system with reduced interference, comprising: a first immunosensor that generates an electrochemical signal based on the formation of a sandwich between an immobilized antibody, a target analyte and a labeled antibody, wherein a portion of the signal arises from non-specific binding of the labeled antibody in the region of the first immunosensor, and

• • a second immunosensor that acts as an immuno-reference sensor and generates a signal that is the same as or predictably related to the degree of non-specific binding which occurs in the region of the first immunosensor, and has an immunocomplex between an immobilized antibody and an endogenous or exogenous protein that is in the sample and that is not the target analyte.

Claims

1. A method for assaying a target analyte while reducing interference in an immunosensor system, comprising:

contacting a sample containing a target analyte plus an endogenous or exogenous protein, which is not the target analyte, with an immunosensor system comprising

a first immunosensor that generates a signal based on the formation of a sandwich between an immobilized antibody, a target analyte and a labeled antibody, wherein a portion of the signal arises from a degree of non-specific binding of the labeled antibody in the region of the first immunosensor,

a second immunosensor that acts as an immuno-reference sensor and generates a signal that is the same as or predictably related to the degree of non-specific binding which occurs in the region of the first immunosensor, and has an immunocomplex between an immobilized antibody and said endogenous or exogenous protein,

washing said first and second immunosensors with a wash fluid, and

using a subtraction analyzer, the signal from the first immunosensor, and the signal from the second immunosensor to determine a corrected analyte concentration in the sample.

2. The method of claim 1, wherein the first and second immunosensors are electrochemical sensors.

3. The method of claim 1, wherein the first and second immunosensors are amperometric electrochemical sensors.

4. The method of claim 1, wherein the first and second immunosensors are selected from the group consisting of potentiometric sensors, field effect transistor sensors, conductimetric sensors, optical sensors, evanescent wave sensors, optical wave guides, thermometric sensors and acoustic wave sensors.

5. The method of claim 1, wherein the system is in a disposable cartridge for measuring analytes in a sample.

6. The method of claim 1, wherein the target analyte is selected from the group consisting of troponin I, troponin T, creatine kinase MB, procalcitonin, hCG, NTproBNP, proBNP, BNP and myoglobin.

7. The method of claim 1, wherein the immobilized antibody in the second immunosensor is to a plasma protein.

8. The method of claim 1, wherein the immobilized antibody in the second immunosensor is to a protein selected from the group consisting of human serum albumin (HAS), bovine serum albumin (BSA), fibrinogen and IgG fc region.

9. The method of claim 1, wherein the endogenous or exogenous protein in the sample is present at a concentration sufficient to bind more than 50% of the available immobilized antibody on the second immunosensor within about 100 seconds of the sample contacting the immunosensor system.

10. The method of claim 1, wherein the immobilized antibody in the second immunosensor has an affinity constant of about 1×

10<sup>−

7</sup> to 1×

10<sup>−

15</sup>.

11. The method of claim 1, wherein the endogenous protein is HAS in a concentration of about 100 ng/ml or more.

12. The method of claim 1, wherein the endogenous or exogenous protein is present in a blood sample at a concentration of at least three orders of magnitude above the affinity constant of the antibody in the second immunosensor.

13. The method of claim 1, wherein both antibodies are immobilized on microparticles of diameter in the range 0.01 μ

m-5.0 μ

m.

14. The method of claim 13, wherein the microparticle is a carboxylate modified polystyrene bead.

15. The method of claim 1, wherein the signals from the first and second immunosensors are used to monitor the washing efficiency.

16. The method of claim 1, wherein the signals from the first and second immunosensors are used to detect anomalous sample conditions including improperly anti-coagulated samples.