Assay device

• US 6,017,767 A
• Filed: 06/05/1995
• Issued: 01/25/2000
• Est. Priority Date: 05/29/1991
• Status: Expired due to Term

• Current Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Original Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Inventors: Chandler, Howard M.
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): NGUYEN, BAO THUY L
• Application Number:
  US08/465,428
• Time in Patent Office: 1,695 Days
• Field of Search: 422/56, 422/57, 422/58, 422/61, 422/99, 435/7.1, 435/7.2, 435/7.9, 435/7.93, 435/7.94, 435/7.95, 435/287.7, 435/287.9, 435/288.4, 435/288.5, 435/969, 435/970, 435/973, 436/518, 436/514, 436/524, 436/807, 436/809, 436/810
• US Class Current: 436/514
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01L 2300/0825 Test strips
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 3/5023 with a sample being transported to, and subsequently stored in an absorbent for analysis
• CPC Class Codes: B01L 3/5055 Hinged, e.g. opposable surfaces
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/30 of temperature
• CPC Class Codes: G01N 30/90 Plate chromatography, e.g. thin layer or paper chromatography
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• 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 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/969 Multiple layering of reactants
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/81 : Tube, bottle, or dipstick

Abstract

A chromatographic assay device for use with immunoassays allows rapid and convenient assays of analytes of biological interest, and permits extractions to be carried out in situ, avoiding the use of separate extraction vessels. The device has a wide dynamic range and avoids interference from particulates or colored components. In one form, the device comprises: (1) a first opposable component comprising a sample preparation means adapted to receive a sample to be assayed; and (2) a second opposable component comprising a chromatographic medium. The first and second opposable components can be brought into opposition so as to cause the sample preparation means to apply the sample to be tested to the chromatographic medium. Preferably, the analyte is detected with a visually detectable label. Other variations of the device vary the arrangement of components to provide optimal chromatography for a variety of analytes, as well as to permit bidirectional chromatography. The devices can be incorporated in test kits, and assay methods using the devices are also disclosed.

Claims

1. A chromatographic assay device for detection or determination of an analyte comprising:

(a) an opposable component including a sample preparation means for receiving a sample to be assayed;

(b) a second opposable component including a chromatographic medium;

wherein the first and second opposable components are brought into opposition so as to cause the sample preparation means to apply the sample to be tested to the chromatographic medium so that the analyte is detected by a labeled second specific binding partner bound to the analyte, the analyte in turn being directly bound to a first specific binding partner bound to the chromatographic medium so that the labeled second specific binding partner is indirectly bound to the first specific binding partner through the analyte, the labeled second specific binding partner either being present in the assay device in either the first or second opposable components prior to the initiation of an assay or being added to the assay device during the performance of the assay.

2. A chromatographic assay device for detection or determination of an analyte comprising:

(a) a first opposable component including a sample preparation means for receiving a sample to be assayed; and

(b) a second opposable component including a chromatographic medium, the chromatographic medium further including a detection zone smaller than the chromatographic medium;

wherein the first and second opposable components are brought into opposition so as to cause the sample preparation means to apply the sample to be tested to the chromatographic medium;

wherein the detection zone contains a first specific binding partner to the analyte immobilized therein so that the analyte is detected by a labeled second specific binding partner to be bound to the analyte, the analyte being in turn directly bound to the first specific binding partner, the labeled second specific binding partner being indirectly bound to the first specific binding partner through the analyte, the labeled second specific binding partner either being present in either the first or second opposable component of the device prior to the initiation of an assay or being added to the assay device during the performance of the assay.

3. The chromatographic assay device of claim 2 wherein the analyte is an antigen or hapten and the first specific binding partner is an antibody to the antigen or hapten.

4. The chromatographic assay device of claim 2 wherein the analyte is an antibody and the first specific binding partner is a hapten or antigen bound specifically by the antibody.

5. A chromatographic assay device for detection or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends;

(ii) a conducting means in operable contact with the first end of the chromatographic medium; and

(iii) an absorbing means in operable contact with the second end of the chromatographic medium; and

(b) a second opposable component including;

(i) a first application means that is a sample application pad; and

(ii) a second application means that is a detector application pad to which detecting reagent is applied;

the first and second application means being positioned on the second opposable component such that they are not in operable contact when the first and second opposable components are not in opposition;

wherein bringing the first and second opposable components into opposition places the conducting means in operable contact with the first application means and places the conducting means in operable contact with the second application means, thereby placing the first and second application means in operable contact with each other and applying the contents of the sample application pad and the detector application pad to the conducting means, and wherein the conducting means transfers the contents of the sample application pad and the detector application pad to the first end of the chromatographic medium of the first opposable component;

wherein the detector application pad contains a first specific binding partner to the analyte in a form that is resolubilized by addition of an aqueous liquid to the detector application pad, the first specific binding partner being labeled with a detectable label, and the chromatographic medium further comprises a detection zone smaller in area than the chromatographic medium, the detection zone containing a second specific binding partner to the analyte immobilized thereto, such that a ternary complex comprising the first specific binding partner, the analyte, and the second specific binding partner forms at the detection zone if analyte is present in the sample, the second specific binding partner being bound directly to the analyte and the labeled first specific binding partner being bound indirectly to the second specific binding partner through the analyte.

6. The chromatographic assay device of claim 5 wherein the detectable label is a visually detectable label.

7. A test kit for the detection or determination of an analyte comprising:

(a) the chromatographic assay device of claim 5; and

(b) an aqueous liquid for resolubilizing the specific binding partner for the analyte labeled with a detectable label, to be applied to the detector application pad.

8. A method of detection or determination of an analyte in a test sample comprising the steps of:

(a) applying the sample to the first application means of the chromatographic assay device of claim 5;

(b) bringing the first and second opposable components of the chromatographic assay device into opposition, such that the sample comprises the aqueous liquid resolubilizing the specific binding partner in the detector application pad, and such that the sample and the resolubilized labeled specific binding partner are applied to the first end of the chromatographic medium;

(c) allowing the sample and the labeled specific binding partner to move through at least a portion of the chromatographic medium, the portion including the detection zone, so that the detection reagent gives a detectable indication of the presence or quantity of the analyte; and

(d) observing or measuring the detection reagent at the detection zone in order to detect or determine the analyte.

9. A chromatographic assay device for the detection or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends;

(ii) a conducting means in operable contact with the first end of the chromatographic medium;

(iii) an absorbing means in operable contact with the second end of the chromatographic medium; and

(iv) a detector application pad in direct contact with the conducting means and positioned such that it is in indirect contact with the first end of the chromatographic medium; and

(b) a second opposable component that is a sample application pad;

wherein bringing the first and second opposable components into opposition causes the sample application pad to apply a sample to be tested to the detector application pad and thus to the first end of the chromatographic medium through the conducting means;

wherein the detector application pad contains a first specific binding partner to the analyte in a form that is resolubilized by addition of an aqueous liquid to the detector application pad, the first binding partner being labeled with a detectable label, and the chromatographic medium further includes a detection zone smaller than the chromatographic medium, the detection zone containing a specific binding partner to the analyte immobilized thereto, such that a try complex comprising the first specific binding partner, the analyte, and the second specific binding partner forms at the detection zone if analyte is present in the sample, the labeled first specific binding partner being indirectly bound to the immobilized second specific binding partner through the analyte in the ternary complex.

10. The chromatographic assay device of claim 9 wherein the contents of the sample application pad after a sample is applied thereto comprise an aqueous liquid, and the aqueous liquid applied to the detector application pad comprises the contents of the sample application pad.

11. A test kit for the detection or determination of an analyte comprising:

(a) the chromatographic assay device of claim 9; and

(b) a specific binding partner for the analyte labelled with a detectable label, to be applied to the second application means.

12. A method for the detection or determination of an analyte in a test sample comprising the steps of:

(a) applying the sample to the sample application pad of the chromatographic assay device of claim 9;

(b) bringing the first and second opposable components of the chromatographic assay device into opposition, such that the sample comprises the aqueous liquid resolubilizing the specific binding partner in the detector application pad and such that the sample and the resolubilized labeled specific binding partner are applied to the first end of the chromatographic medium;

(c) allowing the sample and the labeled specific binding partner to move through at least a portion of the chromatographic medium, the portion including the detection zone, so that the labeled specific binding partner gives a detectable indication of the presence or quantity of the analyte; and

(d) observing or measuring the labeled specific binding partner at the detection zone in order to detect or determine the analyte.

13. A chromatographic assay device for the detection or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends;

(ii) an absorbing means in operable contact with the second end of the chromatographic medium; and

(iii) a detector application pad in direct contact with the first end of the chromatographic medium;

(b) a second opposable component including a sample application pad;

wherein, where the first and second opposable components are brought into opposition, the detector application pad and the sample application pad are in contact except for a region of the sample application pad directly adjacent to the first end of the chromatographic medium, and whereby bringing the first and second opposable components into opposition causes the sample application pad to apply the sample to be tested to the detector application pad and thus the first end of the chromatographic medium;

wherein the detector application pad contains a first specific binding partner to the analyte in a form that is resolubilized by addition of an aqueous liquid to the detector application, the first specific binding partner being labeled with a detectable label, and the detection zone is smaller in area than the chromatographic medium, the detection zone containing a second specific binding partner to the analyte immobilize thereto, such that a ternary complex comprising the first specific binding partner, the analyte, and the second specific binding partner forms at the detection zone if the analyte is present in the sample.

14. The chromatographic assay device of claim 13 wherein the contents of the sample application pad after a sample is applied thereto comprise an aqueous liquid, and the aqueous liquid applied to the detector application pad comprises the contents of the sample application pad.

15. A method for the detection or determination of an analyte in a test sample comprising the steps of:

(a) applying the sample to the sample application pad of the chromatographic assay device of claim 14;

(b) bringing the first and second opposable components of the chromatographic assay device into opposition, such that the sample comprises the aqueous liquid resolubilizing the specific binding partner in the detector application pad, and such that the sample and the resolubilized labelled specific partner are applied to the first end of the chromatographic medium;

(c) allowing the sample and the labelled specific binding partner to move through at least a portion of the chromatographic medium, the portion including the detection zone, so that the labeled specific binding partner gives a detectable indication of the presence or quantity of the analyte; and

(d) observing or measuring the labeled specific binding partner at the detection zone in order to detect the analyte.