Solid phase analytical device and method for using same

• US 5,149,622 A
• Filed: 05/21/1990
• Issued: 09/22/1992
• Est. Priority Date: 10/04/1985
• Status: Expired due to Fees

• Current Assignee: Inverness Medical Switzerland GmbH (Abbott Laboratories)
• Original Assignee: Abbott Laboratories
• Inventors: Clemens, John M., Brown, William E., Knigge, Kevin M., Hofler, John G., Safford, Sarah E., Devereaux, Sharon M.
• Primary Examiner(s): Kepplinger, Esther L.
• Assistant Examiner(s): Spiegel, Carol A.
• Application Number:
  US07/528,277
• Time in Patent Office: 855 Days
• Field of Search: 435/7.1, 435/34, 435/36, 435/288, 435/5, 435/6, 435/805, 435/810, 435/7.92, 435/734, 435/970, 435/971, 436/807-809, 436/527, 436/528, 436/530, 436/531, 436/535, 436/518, 422/55-61
• US Class Current: 435/5
• CPC Class Codes: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: G01N 33/523 the element being adapted for a specific analyte
• CPC Class Codes: G01N 33/528 Atypical element structures, e.g. gloves, rods, tampons, toilet paper
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/54391 based on vertical flow
• CPC Class Codes: G01N 33/56911 Bacteria
• CPC Class Codes: G01N 33/5764 Hepatitis B surface antigen
• CPC Class Codes: G01N 33/76 Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/971 Capture of complex after antigen-antibody reaction

Abstract

A novel material and device useful in solid-phase binding assays to determine the presence or amount of an analyte in a test sample, particularly antigens or antibodies, is disclosed. The material comprises a porous matrix of fibers and a plurality of substantially spherical, solid particles having an average diameter of from about 0.1 to about 5 microns. The particles are retained and immobilized upon the fibers of the matrix. Preferably, the particles have on their surfaces a substance capable of reaction with the analyte in the sample, and the average diameter of the particles is less than the average pore size of the matrix. The device, in a preferred embodiment, comprises a substantially planar layer of the described material.

Claims

1. A method for determining the presence or amount of an analyte in a fluid, comprising the steps of:

a) incubating a sample of the fluid with a plurality of substantially spherical, solid particles having an average diameter of from about 0.1 to about 5 microns, the particles having immobilized upon their surfaces a binding substance which specifically binds the analyte, whereby the analyte becomes bound to the substance to form an analyte/binding substance complex on the particles;

b) contacting a porous, fibrous matrix with the incubated particles, wherein the average diameter of said particles is less than the particle retention rating of said matrix and whereby at least a portion of the incubated particles become retained and immobilized within the matrix upon at least a portion of the fibers thereof;

c) contacting said fibrous matrix containing the complex of the particles with a with a labeled substance capable of specifically binding the analyte, said labeled substance capable of producing a detectable response in the presence of the analyte and an indicator substance, whereby the labeled substance becomes bound to the complex on the particles;

d) washing unbound labeled substance from said fibrous matrix;

e) contacting said fibrous matrix with the indicator substance; and

f) detecting the response produced as a function of the presence or amount of analyte in the sample.

2. The method of claim 1 wherein the labelled substance comprises an enzyme, the indicator substance comprises a chromogen, and detecting the response produced comprises detecting the presence or amount of chromogen in the fibrous matrix.

3. The method of claim 1 wherein the analyte is beta hCG.

4. The method of claim 1 wherein the analyte is a strep A antigen.

5. The method of claim 1 wherein the analyte is HBsAg.

6. A method of determining the presence or amount of an analyte in a test sample, comprising the steps of:

a) contacting a porous matrix of fibers with the test sample wherein substantially spherical, solid particles having a substance adsorbed or bound to their surfaces which specifically binds the analyte, said coated particles having an average diameter between about 0.1 and about 5 microns and wherein the average diameter of said particles is less than the particle retention rating of the matrix are retained and immobilized upon the fibers of said matrix,

b) contacting the porous fibrous matrix of step a) with a second substance which specifically binds the analyte and which is labeled with a substance capable of producing a detectable response in the presence of an indicator substance such that this second substance will bind to any analyte which has bound to the retained and immobilized particles,

c) removing any second substance which has not bound to analyte bound to the retained and immobilized particles,

d) contacting the porous matrix of step of step c with the indicator substance, and

e) correlating any detectable response produced to the presence or amount of the analyte in the sample.

7. The method of claim 6, wherein the analyte is beta-hCG.

8. The method of claim 6, wherein the analyte is Strep-A antigen.

9. The method of claim 6, wherein the analyte is HBsAg.

10. The method of claim 6 wherein:

the detectable response resulting from the interaction of the second substance and the indicator substance is a visually detectable response, and prior to step a)

a second set of substantially spherical particles are retained and immobilized in the matrix, said second set of particles having a substance adsorbed or bound to their surfaces which specifically binds said second substance and having an average diameter between about 0.1 and about 5 microns and wherein the average diameter of said second particles is less than the particle retention rating of the matrix,

the particles with an analyte binding substance adsorbed or bound to their surfaces and said second set of particles are arranged in a pattern such that after the matrix is contacted with the sample, the second substance and the indicator substance;

i) a visually detectable plus sign is displayed if detectable analyte is present in the sample as a result of indicator substance interacting with both second substance adhered to said second set of particles and second substance adhered to analyte adhered to said analyte binding particles and

ii) a visually detectable minus sign is displayed if no detectable analyte is present in the sample as a result of indicator substance interacting with only the second substance adhered to the second set of particles.

11. A solid-phase assay device useful in a binding assay to determine the presence or an amount of an analyte in a fluid sample comprising

a) a rigid carrier means for holding a porous matrix of fibers,

b) a porous matrix of fibers arranged in a substantially planar layer having a first sample-contacting surface and a second surface opposed to said first surface, said matrix being disposed in said carrier means such that, when the device is used in the performance of an assay, at least a portion of the sample contacting the first surface passes through said substantially planar layer to the second surface, and

c) substantially spherical, solid particles retained and immobilized on the fibers of said porous matrix, said particles having a substance adsorbed or bound to their surfaces which specifically binds analyte, said coated particles having an average diameter between about 0.1 and about 5 microns and wherein the average diameter of said particles is less than the particle retention rating of said porous matrix.

12. The solid phase assay device of claim 11, wherein the device additionally comprises filtering means disposed in relationship to the first surface of said substantially planar layer, such that, when the device is in use, sample fluid passes through said filtering means prior to contacting the first surface.

13. The solid phase assay device of claim 11, wherein the device additionally comprises absorbent means disposed in relationship to the second surface of said substantially planar layer, such that at least a portion of sample passing through said substantially planar layer is absorbed by said absorbent means.

14. The solid phase assay device of claim 13, wherein the device additionally comprises a barrier layer interposed between said substantially planar layer and said absorbent means, said barrier layer having one way permeability such that when the device is in use and a fluid is passed through said substantially planar layer, it restricts fluid retained by said absorbent means from returning to said substantially planar layer.

15. The solid phase assay device of claim 14, wherein the barrier layer is composed of a polyethylene weave material.

16. In a binding assay utilizing a solid-phase to determine the presence or amount of an analyte in a test sample comprising contacting the test sample with an immunoreactive porous matrix of fibers in order to capture the analyte and detecting the captured analyte, the improvement comprising capturing the analyte in a porous matrix of fibers in which substantially spherical, solid particles are retained and immobilized, said particles having a substance adsorbed or bound to their surfaces which specifically binds the analyte, said coated particles having an average diameter between about 0.1 and about 5 microns and wherein the average diameter of said particles is less than the particle retention rating of the porous matrix.

17. The binding assay of claim 16, wherein the binding assay is an enzyme immunoassay.

18. The binding assay of claim 16, wherein the binding assay is a competitive binding assay.