SIMULTANEOUS MULTIANALYTE ELECTROCHEMICAL ASSAY BASED ON SPATIAL RESOLUTION

• US 20010029048A1
• Filed: 03/12/1999
• Published: 10/11/2001
• Est. Priority Date: 03/16/1998
• Status: Active Grant

• Current Assignee: University of Cincinnati
• Original Assignee: University of Cincinnati
• Inventors: DING, YING, HEINEMAN, WILLIAM R., HALSALL, BRIAN
• Granted Patent: US 7,297,312 B2
• Time in Patent Office: Days
• US Class Current: 436/518
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: Y10S 435/971 Capture of complex after antigen-antibody reaction
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte

Abstract

The present invention is premised on the realization that a single electrochemical binding assay device can be used to determine multiple analytes in a single sample by simultaneous amperometric measurements using a plurality of working electrodes.

More particularly the present invention is premised upon the realization that by establishing different analyte binding sites i.e., antibodies or antigens on a solid phase at a distinct separate location and locating separate working electrodes within proximity of those separate locations, one can add enzyme labeled antibodies or antigens depending on the assay format, and then quantitate the amount of enzyme reaction product, whether chemically the same or different, generated by simultaneous amperometric measurement with the independent micro electrode for each area. The independent micro electrode for each area is spatially separated from adjacent analyte so that a measurement can be taken before cross-interference due to diffusion of product from adjacent analyte areas. The objects and advantages of the present invention will be further appreciated in light of the following detailed descriptions and drawings in which:

Claims

1. A simultaneous electrochemical assay comprising a substrate having a plurality of analyte binding areas, each of said analyte binding areas adapted to bind a different specific analyte;

and a plurality of working electrodes each working electrode adjacent to one analyte binding area and separated from the nearest adjacent analyte binding area by a distance effective to permit a measurement by each electrode of analyte bound to its adjacent analyte binding site and not measure an interfering amount of analyte bound to said nearest adjacent analyte binding site.

2. The assay claimed in

claim 1

wherein said binding sites comprise a plurality of different analyte specific proteins.

3. The assay claimed in

claim 1

wherein said binding sites each comprise a different antigen.

4. The assay claimed in

claim 1

wherein each binding site comprises a different antibody.

5. The assay claimed in

claim 1

further comprising at least one auxiliary electrode.

6. A method of testing for a plurality of different analytes in a test solution using a test cell having a plurality of spaced analyte binding sites wherein each binding site is specific for a separate analyte;

locating separate electrodes adjacent to each binding site and spaced from an adjacent binding site;

adding a test solution to said cell;

adding reagent to said cell wherein portions of said reagent react with each of said analytes and wherein said reagent includes at least one label

electrochemically detecting said label at each of said electrodes in less than a time in which label-produced product at any binding site can migrate to an adjacent binding site.

7. The method claimed in

claim 6

wherein said label contains an enzyme and further comprising adding substrate to said cell wherein said label is detected by measuring a reaction product of said enzyme and said substrate.

8. The method claimed in

claim 7

wherein said product is measured amperometrically.

9. The method claimed in

claim 6

wherein said binding site comprises a plurality of analyte specific proteins and said reagent comprises a plurality of analyte specific proteins each labeled with the same label.