# US 2010/0167264 A1 · Quantitative analyte assay device and method

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- Number: US 2010/0167264 A1
- Document ID: US20100167264A1
- Status: Published application
- Kind: A1
- Application number: 12/317,814
- Filed: 2008-12-30
- Published: 2010-07-01
- Earliest claimed: 2008-12-30
- Inventors: Jin Po Lee
- Web page: https://patentgrep.com/patent/US20100167264A1

## CPC

- G01N 33/54388: {based on lateral flow}
- G01N 21/78: producing a change of colour
- G01N 33/74: involving hormones {or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors}
- G01N 2021/6439: {with indicators, stains, dyes, tags, labels, marks}
- G01N 2021/7786: {Fluorescence}
- G01N 2021/7793: {Sensor comprising plural indicators}
- G01N 2021/7796: {Special mountings, packaging of indicators}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The present invention relates to an assay device and a method for using such for the quantitative determination of an analyte, based on a test strip, which contains a porous test membrane allowing for capillary flow of the analyte and complexes of the analyte, a porous upstream membrane in fluid connection with the test membrane and a porous downstream membrane in fluid connection with the test membrane, wherein the test membrane contains a test site having immobilized thereon a ligand capable of reacting with the analyte and binding such to the test site, and two standard band sites having immobilized thereon known high and low concentrations of a calibrator agent capable of reacting with a label conjugate and binding such to the standard sites, wherein the upstream membrane has a site for the application of a sample to be analyzed, and has a site downstream from the sample application site for depositing label conjugates capable of reacting with the analyte and label conjugates capable of reacting with the immobilized calibrator agents in the standard bands to provide a known label response in the standards bands, and the downstream membrane is capable of absorbing said sample and providing the capillary flow for the sample through the upstream and test membrane.

## Claims

### Claim 1

An assay device for the quantitative determination of an analyte, comprising a test strip containing a porous test membrane allowing for capillary flow of the analyte and complexes of the analyte, a porous upstream membrane in fluid connection with the test membrane and a porous downstream membrane in fluid connection with the test membrane,
(a) said test membrane containing a test site having immobilized thereon a ligand capable of reacting with the analyte and binding such to the test site, and two standard band sites having immobilized thereon known high and low concentrations of a calibrator agent capable of reacting with a label conjugate and binding such to the standard sites,
(b) said upstream membrane having a site for the application of a sample to be analyzed, and having a site downstream from the sample application site for depositing label conjugates capable of reacting with the analyte and label conjugates capable of reacting with the immobilized calibrator agents in the standard bands to provide a known label response in the standards bands, and
(c) said downstream membrane capable of absorbing said sample and providing the capillary flow for the sample through the upstream and test membrane.

### Claim 2 (depends on 1)

The assay device of claim 1 wherein the label conjugates on the upstream membrane give rise to a color.

### Claim 3 (depends on 2)

The label conjugates of claim 2 comprising gold conjugates, fluorescent dye conjugates, latex conjugates and enzyme conjugates.

### Claim 4 (depends on 1)

The device of claim 1 wherein the test strip is contained in a cassette allowing for the addition of the sample at the upstream membrane and viewing the results of color formation at the sites of the test membrane.

### Claim 5 (depends on 1)

The device of claim 1 wherein the label conjugates are different for the analyte and the calibrator agents.

### Claim 6 (depends on 6)

The device of claim 6 wherein the calibrator agents and corresponding label conjugates are different for the high and low calibrator agent concentration.

### Claim 7 (depends on 1)

The device of claim 1 wherein the calibrator agents are the same as the analyte and are capable of binding to the label conjugate for the analyte.

### Claim 8 (depends on 1)

The method of quantitative analysis of an analyte in a sample comprising
(a) establishing the label response of known amounts of the calibrator agents on the test strip of the device of claim 1 and label conjugate capable of binding to the calibrator agents
(b) establishing the label response of known quantities of an analyte label conjugate when bound to the immobilized ligand in the test band of the device of claim 1,
(c) correlating the label responses of the calibrator bands with the label responses of known concentrations of analyte label conjugates in the test band
(d) adding a sample suspected of containing an analyte for which the test strip of claim 1 was designed to the device of claim 1
(e) measuring the label response at the test zone and comparing such to the standard curve of step (c) for the label responses in the standard bands as established in step (d), and
(e) converting the measured label responses to concentration of analyte in the sample

### Claim 9 (depends on 8)

The method of claim 8 wherein the label generates a visual signal at the binding sites of the assay device.

### Claim 10 (depends on 9)

The method of claim 9 wherein the visual signal is generated by a colloidal gold conjugate, a fluorescent dye conjugate, a latex particle conjugate or an enzyme conjugate.

### Claim 11 (depends on 8)

The method of claim 8 wherein the analyte is an antigen associated with an infectious agent such as a virus, bacterium fungus or a prion.

### Claim 12 (depends on 8)

The method of claim 8 wherein the analyte is a hormone such as hCG, thyroxin, TSH, glucagons, insulin relaxin, prolactin, melanotropin, gastrin, somatotropin, gastrin and bradykin.

### Claim 13 (depends on 8)

The method of claim 8 wherein the analyte is a cancer or tumor marker such as prostrate specific antigen (PSA), carcinoembryonic antigen (CEA), and alpha-fetoprotein.

### Claim 14 (depends on 8)

The method of claim 8 wherein the analyte is a cardiac marker.

### Claim 15 (depends on 8)

The method of claim 8 wherein the calibrator agents are monoclonal ligands.

### Claim 16 (depends on 15)

The method of claim 15 wherein the calibrator agents are BTG antibodies, rabbit IgG antibodies and the label conjugates are label bound antigens capable of binding to the calibrator agents.