Reagents for reducing leukocyte interference in immunoassays

Patent metadata

Field	Value
Publication number	US-9316638-B2
Application number	US-201514673364-A
Country	US
Kind code	B2
Filing date	Mar 30, 2015
Priority date	Apr 30, 2010
Publication date	Apr 19, 2016
Grant date	Apr 19, 2016

Abstract

Methods and devices for reducing interference from leukocytes in an analyte immunoassay are provided. In one embodiment, a method is provided comprising the steps of amending a biological sample such as a whole blood sample with one or more leukocidal reagents that reduce or eliminate the metabolic activity of leukocytes, and performing an immunoassay on the amended sample to determine the concentration of analyte in the sample. Preferably, the sample is amended with one or more enzymes and optionally one or more enzyme substrates and cofactors.

First claim

What is claimed is:

1. A kit for performing an electrochemical immunoassay for a target analyte in a whole blood sample comprising:
a leukocidal reagent configured to substantially inhibit phagocytic activity of leukocytes in the whole blood sample, wherein the leukocidal reagent is a mitochondrial electron transport inhibitor or a mitochondrial membrane uncoupling agent, and
electrochemical immunoassay reagents for detecting at an immunosensor the target analyte in the leukocidal reagent-treated whole blood sample.

2. The kit of claim 1 , wherein the leukocidal reagent is the mitochondrial electron transport inhibitor.

3. The kit of claim 1 , wherein the leukocidal reagent is the mitochondrial membrane uncoupling agent.

4. The kit of claim 1 , wherein the leukocidal reagent is the mitochondrial electron transport inhibitor or the mitochondrial membrane uncoupling agent and selected from the group consisting of cyanide, antimycin, rotenone, malonate, carbonyl cyanide p-[trifluoromethoxyl]-phenyl-hydrozone, 2,4-dinitrophenol, and oligomycin.

5. The kit of claim 1 , further comprising the immunosensor comprising an immobilized first antibody to the target analyte, and a labeled second antibody to the target analyte, wherein the immunoassay is a sandwich assay.

6. The kit of claim 1 , further comprising the immunosensor comprising an immobilized antibody to the target analyte, and the target analyte is labeled, wherein the immunoassay is a competitive assay.

7. The kit of claim 1 , further comprising beads opsonized for the leukocytes.

8. The kit of claim 1 , wherein the whole blood sample is amended with an anticoagulant.

9. The kit of claim 1 , wherein the target analyte is selected from the group consisting of Troponin I (TnI), Troponin T (TnT), Brain Natriuretic Peptide (BNP), N-terminal of the Prohormone Brain Natriuretic Peptide (NTproBNP), Prohormone Brain Natriuretic Peptide (proBNP), Human Chorionic Gonadotropin (HCG), Thyroid-Stimulating Hormone (TSH), Neutrophil Gelatinase Associated Lipocalin (NGAL), theophylline, digoxin, and phenytoin.

10. The kit of claim 1 , further comprising the immunosensor.

11. The kit of claim 10 , wherein the leukocidal reagent is positioned upstream of the immunosensor such that the whole blood sample is contacted with the leukocidal reagent prior to contacting the immunosensor.

Assignees

• Abbott Point Of Care Inc
• Abbott Point Of Care Inc

Inventors

• Campbell John Lewis Emerson
• Davis Graham
• Omakor John Emegbero
• Moss Adam Roger
• Guhados Ganesh

Classifications

• Y10T436/25375

  Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.] · CPC title

• Y10T436/25125

  Digestion or removing interfering materials · CPC title

• G01N33/5375

  by changing the physical or chemical properties of the medium or immunochemicals, e.g. temperature, density, pH, partitioning · CPC title

• G01N33/54393 Primary

  Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding · CPC title

• G01N33/5306 Primary

  Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding · CPC title