QUANTITIVE ANALYTE ASSAY DEVICE AND METHOD

• US 20170199182A1
• Filed: 01/19/2017
• Published: 07/13/2017
• Est. Priority Date: 12/30/2008
• Status: Abandoned Application

• Current Assignee: Assurance, LLC
• Original Assignee: Assurance, LLC
• Inventors: LEE, Jin Po
• Application Number:
  US15/410,424
• Publication Number:
  US 20170199182A1
• Time in Patent Office: Days
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 2021/7793 Sensor comprising plural indicators
• CPC Class Codes: G01N 2021/7796 Special mountings, packaging of indicators
• CPC Class Codes: G01N 21/78 producing a change of colour
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• 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: G01N 33/57434 of prostate
• CPC Class Codes: G01N 33/57473 involving carcinoembryonic antigen, i.e. CEA
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
• CPC Class Codes: G01N 33/78 Thyroid gland hormones , e.g. T3, T4, TBH, TBG or their receptors
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10S 436/813 Cancer
• CPC Class Codes: Y10S 436/814 : Pregnancy
• CPC Class Codes: Y10S 436/818 : Human chorionic gonadotropin

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

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 at least two standard band sites having immobilized thereon known high and low concentrations of at least two different calibrator agents capable of reacting with a label conjugated antibody 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 removably depositing label conjugates capable of reacting with the analyte and at least two 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.

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

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

4. The device of claim 1 wherein the test membrane contains two calibrator agents.

5. The device of claim 4 wherein the calibrator agents are rabbit IgG and bovine thyroid globulin.

7. 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.

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

10. The device of claim 1 wherein the test membrane contains three standard bands.

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

14. 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.

15. 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.

16. 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.

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

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

19. 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.

20. The method of claim of claim 8 where more than two calibrator agents are used.

6.

9. The device of claim 6 wherein the test membrane is microporous nitrocellulose and the upstream membrane is fiberglass.

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

11. The method for the quantitative analysis of an analyte in a sample comprising

(a) immobilizing on a microporous test strip in a test band an antibody to the analyte to be established in concentrations adequate to bind to analyte concentrations in the sample,

(b) immobilizing different calibrator agents on the test strip in at least two standard bands in the amounts necessary which when treated with excess of labeled antibodies to the immobilized calibrator agent result in a measurable response corresponding to different concentration of analyte,

(c) removably depositing on an additional upstream membrane in contact with the test strip labeled antibodies to the analyte and the calibrator agents

(d) establishing the curve showing the measurable conjugate response to different analyte concentration,

(e) adding a sample suspected of containing the analyte for which the test strip was designed to the assay strip and complexing such to an antibody conjugated to a measurable label and adding measurable labeled antibodies to the calibrator agents to the sample prior to contact with the standard and test bands,

(f) measuring the label response at the test and standard bands and comparing such to the standard curve of step (d) for the label responses in the standard bands as established in step (b) and

(g) converting the measured label responses to concentration of analyte in the sample.