System for electrochemical quantitative analysis of analytes within a solid phase

• US 6,713,308 B1
• Filed: 05/12/2000
• Issued: 03/30/2004
• Est. Priority Date: 05/05/1999
• Status: Expired due to Fees

• Current Assignee: InTec Science, Inc.
• Original Assignee: InTec Science, Inc.
• Inventors: Lu, Fang, Lu, Frank N. W., Wang, Kai Hua
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Cook, Lisa V.
• Application Number:
  US09/701,922
• Time in Patent Office: 1,418 Days
• Field of Search: 422/56-58, 422/70, 422/82.01, 422/82.03, 422/98, 422/99, 422/110, 435/3, 435/6, 435/287.1, 435/7.1, 435/7.93, 435/287.2, 435/805, 435/810, 435/970, 436/164, 436/169, 436/172, 436/514, 436/518, 436/516, 436/528, 436/530, 436/533, 436/534, 436/536, 436/538, 436/541, 436/800, 436/805, 436/807, 436/810, 436/815, 436/829, 436/901, 424/12, 232/30.B, 204/195.B, 204/1.E, 204/195.M, 204/195.L, 204/193, 204/286.1, 204/288
• US Class Current: 436/514
• CPC Class Codes: G01N 2458/30 Electrochemically active labels
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: Y10S 435/97 Test strip or test slide

Abstract

The analytical process utilized in the system of this invention comprises three (3) step distinct steps wherein an analyte of interest in a test sample is initially labeled and subsequently isolated within a porous medium of a test device. Once isolated within the medium, the label is displaced from the analyte, or from the complex with the analyte, and converted, under electrolytic condition, to a metallic species which is caused test to deposit upon a working electrode of the test device. This working electrode is part of an electrode array that is positioned coincident with the porous medium, yet maintained physically remote therefrom. This deposit is then stripped from the working electrode, under anodic stripping conditions, and the current generated within the electrode array monitored. The characteristic response curve that is produced thereby can be correlated with the identity and concentration of the analyte(s) with the test sample.

Claims

1. A system suitable for concurrent electrochemical, quantitative determination of multiple analytes in a common test fluid sample, comprising:

A. at least one test kit reagent comprising a labeled substance that is both specific for interaction with an analyte of interest or mimicking of an analyte of interest within said system, and electrochemically active so as to be capable of electrolytic reduction and oxidation, under analysis conditions;

B. a solid phase, porous, electrochromatographic medium having a common, linear fluid pathway defined therein, said linear pathway having a delimited area containing at least one immobilized binding material specific for binding to at least one labeled substance or to a complex of a labeled substance and an analyte suspected of being present in said common fluid test sample;

C. means for combining said test kit reagent(s) and a common fluid test sample suspected of containing one or more analytes of interest within said solid phase, porous electrochromatographic medium, so as to cause said common fluid sample containing said labeled substance(s) and/or a complex of a labeled substance(s) and one or more of said analytes of interest, to flow along said fluid pathway of said medium to said delimited area of said medium and thereby contact said immobilized binding material within said delimited area of said medium, under binding conditions, whereby said labeled substance(s), and/or a complex of a labeled substance(s) and an analyte of interest, is bound by said immobilized binding material within said delimited area of said medium;

D. electrode means coincident with and in spaced-apart relationship with said delimited area of said medium, said electrode means including at least one working electrode and second electrode selected from the group consisting of a reference electrode and a counter electrode;

E. means for displacement, under electrolytic conditions, of said label(s) from each of said labeled substance(s), and/or from each of said complex of a labeled substance and analyte(s), bound within said delimited area of said medium, so as to cause said label to form a deposit upon said working electrode of said electrode means;

F. means for activation of said working electrode, under stripping conditions, so as to effect oxidation of said deposit and thereby produce a current signal response indicative of the concentration of each said analyte(s) of said common test fluid and

G. means for correlation of said current signal response with a standard or reference.

2. The system of claim 1, wherein the solid phase, porous electrochromatographic medium includes a second delimited area and a second means for establishing electrical contact therewith under assay conditions, and means for comparison or reference thereof to said electrochemical changes, characteristic of each label.

3. The system of claim 1, wherein the labeled substance comprises a metal selected from the group consisting of Mg, Al, Ca, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, Sr, Y, Zr, Mo, Tc, Pd, Cd, In, Sn, Sb, Ba, Hf, W, Re, Hg, Tl, Pb, La, Ce, Rh, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Yd, Th, U, Pu and the isotopes thereof.

4. The system of claim 1, wherein the labeled substance comprises an analyte mimic, a complex of an analyte mimic with a labeled ligand or a liposome, wherein said substance is electrochemically active under electrochemical analytical conditions.

5. The system of claim 1, wherein the electrochemical analytical conditions comprise potentiostatic measurement of an electrochemically active substance indicative of the analyte of interest.

6. The system of claim 1, wherein said means for combining said test kit reagent(s) and a common fluid test sample suspected of containing one or more analytes of interest within said solid phase includes (a) a housing having a top section and base section, wherein at least one of said top section or base section has means for coupling said base to said top section, so as to seal said solid phase with said housing, and thereby align said solid phase relative to a series of apertures in said top section of said housing, and (b) means, associated with an aperture of said housing, for isolating a test site within said solid phase, so as to direct test kit reagents onto said test site and thereby confine interaction of test kit reagents and said fluid sample to within a delimited area of said solid phase.