One-step lateral flow nonbibulous assay

• US 5,766,961 A
• Filed: 11/23/1993
• Issued: 06/16/1998
• Est. Priority Date: 01/11/1991
• Status: Expired due to Term

• Current Assignee: Quidel Corporation
• Original Assignee: Quidel Corporation
• Inventors: Pronovost, Allan D., Pawlak, Catherine, Goins, Keren
• Primary Examiner(s): Woodward, Michael P.
• Application Number:
  US08/156,877
• Time in Patent Office: 1,666 Days
• Field of Search: 422/56, 422/61, 436/510, 436/514, 436/518, 436/520, 436/525, 436/532, 436/533, 436/534, 436/535, 436/807, 436/810, 436/829
• US Class Current: 436/510
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• 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 33/76 Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/814 Pregnancy
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation

Abstract

A particularly efficient design for a nonbibulous lateral flow one step assay for an analyte in a biological sample is disclosed. In the improved device of the invention, three zones which are in nonbibulous lateral flow contact are employed: a sample receiving zone, a labeling zone, and a capture zone. The sample containing analyte is carried through the labeling zone and interacts with an assay label comprising visible moieties, preferably particles, which are coupled to specific binding reagent for analyte or to a competitor with analyte for a capture reagent. The flow continues into the capture zone where the visible moieties to which analyte or competitor are coupled are captured. Excess fluid is absorbed into an absorbent zone in contact with the capture zone. A positive result is obtained by visualizing the visible moieties in the capture zone. An additional improvement comprises obtaining the nonbibulous lateral flow by converting a bibulous support to a nonbibulous support through coating with a blocking agent, such a methylated BSA. Control label comprising visible moieties (preferably, visually distinguishable from those of the assay label) may also be included in the labeling zone and captured in a separate control portion of the capture zone to verify that the flow of liquid is as expected.

Claims

1. An assay device for detection of the presence or absence of an analyte in a sample, said assay device providing lateral flow contact between three separately prepared zones, said zones comprising:

(a) a sample receiving zone prepared by lyophilizing a first matrix through which all dissolved or dispersed components in a liquid sample flow at substantially equal rates, said sample receiving zone being in lateral flow contact with

(b) a labeling zone prepaired by reversibly coupling a label

which label comprises visible moieties coupled either to a specific binding partner reactive with the analyte or to a ligand competitive with analyte for binding to capture reagent

to a second matrix in the presence of a methylated BSA blocking agent so that all dissolved or dispersed components in a liquid sample will flow at substantially equal rates through said second matrix, followed by lyophilizing said second matrix said labeling zone being in lateral flow contact with

(c) a capture zone prepared by irreversibly coupling a capture reagent which binds to analyte to at least a portion of a third matrix followed by treating said third matrix with a methylated BSA blocking agent so that all dissolved or dispersed components in a liquid sample will flow in substantially equal rates said capture zone being in fluid contact with an absorbent;

wherein the presence or absence of analyte in the sample is detectable in the capture zone within one minute of adding the sample to the sample receiving zone.

2. The device of claim 1 wherein said capture reagent is selected from the group consisting of an antibody and an immunologically reactive fragment thereof specific for analyte.

3. The device of claim 1 wherein said assay label comprises visible moieties coupled to a specific binding partner for said analyte so as to bind said analyte to said specific binding partner.

4. The device of claim 3 wherein said specific binding partner for analyte is selected from the group consisting of an antibody and an immunologically reactive fragment thereof.

5. The device of claim 1 wherein said assay label comprises visible moieties coupled to a competitor of said analyte for said capture reagent.

6. The device of claim 5 wherein said competitor of said analyte is additional analyte or similarly immunologically active analog thereof.

7. The device of claim 1 wherein said capture zone matrix is selected from the group consisting of nitrocellulose, nitrocellulose blends with polyester or cellulose, and nylon.

8. The device of claim 1 wherein the labeling zone further includes a control label comprising visible moieties coupled to a control binding reagent, and wherein said capture zone further includes a specific binding partner for said control binding reagent in a portion of said capture zone different from that wherein said capture reagent for said assay label is located.

9. The device of claim 8 wherein said control reagent is avidin or a derivative thereof and wherein said specific binding partner for the control reagent is biotin.

10. The device of claim 8 wherein said assay label and control label are visually distinguishable.

11. The device of claim 10 wherein said assay label and control label are of different colors.

12. The device of claim 1 wherein said visible moieties are particles.

13. The device of claim 12 wherein said particles are selected from the group consisting of colored latex beads, red blood cells, metal sols and liposomes containing dye.

14. A method to determine the presence or absence of analyte in a sample, which method comprises applying said sample to the sample receiving zone of the device of claim 1, so as to permit said sample to flow through the labeling zone and the capture zone into the absorbent zone, and

detecting the presence or absence of visible moieties in the capture zone in said portion thereof containing the capture reagent capable of binding said assay label.

15. The method of claim 14 wherein said analyte is human chorionic gonadotropin (hCG).

16. The method of claim 15 wherein the specific binding partner for analyte is a monoclonal antibody immunoreactive with hCG.

17. The method of claim 16 wherein the capture reagent is rabbit anti-hCG.

18. The method of claim 14 wherein the visible moieties are particles.

19. The method of claim 14 wherein the particles are selected from the group consisting of colored latex beads, red blood cells, metal sols and liposomes containing dye.

20. The device of claim 1, wherein said matrices of said sample receiving zone, labeling zone and capture zone have been converted from a form in which all dissolved or dispersed components in a liquid sample do not flow at substantially equal rates to a form in which all dissolved or dispersed components in a liquid sample flow at substantially equal rates.

21. The method of claim 14, wherein said matrices of said sample receiving zone, labeling zone and capture zone have been converted from a form in which all dissolved or dispersed components in a liquid sample do not flow at substantially equal rates to a form in which all dissolved or dispersed components in a liquid sample flow at substantially equal rates.