Binding assay device

• US 5,604,110 A
• Filed: 06/07/1995
• Issued: 02/18/1997
• Est. Priority Date: 10/30/1985
• Status: Expired due to Term

• Current Assignee: Lonza Group AG
• Original Assignee: Celltech Group Limited (UCB SA)
• Inventors: Fleming, Ian M., Perry, Martin J., Baker, Terence S.
• Primary Examiner(s): Hutzell, Paula K.
• Assistant Examiner(s): Wolski, Susan C.
• Application Number:
  US08/488,080
• Time in Patent Office: 622 Days
• Field of Search: 422/56, 435/7.92, 435/7.93, 435/7.94, 435/28, 435/805, 435/806, 435/962, 435/970, 436/501, 436/514, 436/518, 436/535, 436/810, 436/817
• US Class Current: 435/7.92
• CPC Class Codes: G01N 33/54386 Analytical elements
• 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 435/805 Test papers
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/817 Steroids or hormones

Abstract

A device for performing an enzyme-labelled binding assay comprises an absorbent material and a developing solution, wherein the absorbent material is provided with a plurality of reagent zones including an indicator reagent zone, and is capable of transporting the developing solution by capillary action sequentially through each reagent zone, and wherein the indicator reagent zone includes a reagent capable, directly or indirectly, of immobilizing an enzyme-labelled reagent in an amount dependent upon the assay result, characterized in that the developing solution includes a signal producing substrate for the enzyme. The substrate moves slower through the absorbent material than the enzyme-labelled reagent or any compound of the enzyme-labelled reagent formed in the assay. The absorbent material is suitably in the form of an elongate strip provided with transverse reagent zones. The device is useful form performing immunoassays including immunometric assays and dual analyte assays.

Claims

1. A test system for performing a binding assay for determining the presence or absence of analyte in a sample or the relative concentrations of two analytes in a sample, comprising:

a) an absorbent material in the form of an elongate strip having a sample application zone upstream of a plurality of transverse reagent zones, wherein said reagent zones of said absorbent material comprise, at least an enzyme-labelled reagent zone which includes an enzyme-labelled species comprising an enzyme-labelled analyte or an enzyme-labelled reagent that binds to said analyte, such that in use, said enzyme-labelled species is caused to migrate through the strip by passage of a developing solution through the strip, and an indicator reagent zone which includes an immobilized reagent that, directly or indirectly, binds and thereby immobilizes said enzyme-labelled species in an amount dependent on the quantity of said analyte present in said sample; and

b) a developing solution, wherein said developing solution comprises a signal-producing substrate for the enzyme that generates signal only in or downstream from said indicator reagent zone, wherein the signal-producing substrate is a single color-producing compound or a compound acting as a cofactor with a further compound or compounds to produce a signal in the presence of enzyme, wherein said any further compound or compounds are present in the developing solution;

wherein said developing solution is initially in contact only with that portion of said absorbent material upstream from said indicator zone but in which ultimately, by capillary action, sequentially contacts all reagent zones of said absorbent material;

wherein said enzyme-labelled species is mobilized in said developing solution but does not react with said signal producing substrate to produce color except at or downstream form said indicator reagent zone; and

wherein said signal-producing substrate is transported by the developing solution slower than, said enzyme-labelled species.

2. The test system according to claim 1 wherein the developing solution and the signal-producing substrate for the enzyme are contained in a rupturable sac adjoining the absorbent material.

3. The test system according to claim 1 wherein the enzyme is horseradish peroxidase and the developing solution contains tetramethylbenzidine and hydrogen peroxide.

4. The test sheet comprising a rigid plastic backing plate or stand with a plurality of systems according to claim 1 attached thereto.

5. The test system according to claim 1 wherein the enzyme-labelled reagent and the immobilized reagent are antibodies.

6. The test system according to claim 1 wherein the assay is a dual analyte assay for determining the relative concentrations of pregnanediol-3-glucuronide and oestrone-3-glucuronide in urine.