Direct read lateral flow assay for small analytes

• US 5,798,273 A
• Filed: 09/25/1996
• Issued: 08/25/1998
• Est. Priority Date: 09/25/1996
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co, EMD Millipore Corporation (Merck & Co., Inc.)
• Original Assignee: Becton, Dickinson & Co, Millipore Corporation
• Inventors: Lovell, Stephen J., Fisher, Abigail S., Rosenstein, Robert W., Weiss, Alan J., Shuler, John K.
• Primary Examiner(s): Smith, Lynette F.
• Assistant Examiner(s): NELSON, BRETT L
• Application Number:
  US08/719,221
• Time in Patent Office: 699 Days
• Field of Search: 436/514, 436/501, 436/507, 436/510, 436/512, 436/513, 435/7.5, 435/7.93
• US Class Current: 436/514
• 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

Abstract

The present invention relates to a method and assay device for detecting small analytes. The results of the assay can be directly read from the device, which is a lateral flow device.

Claims

1. A process for determining the presence or amount of an analyte in a sample comprising:

a) contacting said sample with a preparation containing

i) anti-analyte antibodies, which bind to analyte or to both analyte and an analyte analog, and

ii) an assay tracer, wherein said assay tracer comprises a detectable particulate substance having immobilized on its surface, analyte or analyte analog and a first member of a binding pair, to provide a sample/preparation mixture containing analyte/anti-analyte antibody complexes and/or assay tracer/anti-analyte complexes;

b) contacting said sample/preparation mixture with a sample addition area of a lateral flow device, wherein said device comprises a solid support containing, in sequence,

i) said sample addition area,

ii) a capture area, wherein analyte or analyte analog is immobilized, to capture said analyte/anti-analyte antibody complexes and/or assay tracer/anti-analyte complexes,

iii) a read-out area comprising at least one zone, wherein a second member of said binding pair, which binds to said first member, is immobilized, to bind assay tracer not captured in said capture area, in proportion to the amount of analyte in said sample;

c) allowing said sample/preparation mixture to flow through said lateral flow device; and

d) detecting the presence or amount of assay tracer bound in said at least one zone of said read-out area as a measure of analyte in said sample.

2. The process of claim 1, wherein said assay tracer is a colored particle.

3. The process of claim 2, wherein said colored particle is a colored liposome or a colored polymeric bead.

4. The process of claim 3, wherein said colored polymeric bead is a colored polystyrene bead.

5. The process of claim 1, wherein said solid support comprises nitrocellulose.

6. The process of claim 1, wherein said first member of said binding pair, immobilized on said assay tracer, is biotin.

7. The process of claim 1, wherein said second member of said binding pair, immobilized in said read-out zone, is avidin, streptavidin, or neutravidin.

8. The process of claim 1, wherein:

a) said lateral flow device further comprises a housing which covers the entire lateral flow device, wherein said housing has

i) an opening over said sample addition area,

ii) a first window for viewing said read-out area, and

iii) a second window for viewing a distal end of said device; and

b) said process further comprises contacting said sample/preparation mixture with said sample addition area by pouring said sample/preparation mixture through said opening.

9. The process of claim 8, wherein an end-of-assay tracer is added to said sample/preparation mixture for viewing through said second window.

10. The process of claim 1, wherein:

a) said lateral flow device further comprises a second zone in said read-out area, which functions as a control, wherein an anti-control-antigen antibody is immobilized; and

b) said process further comprises contacting a control tracer with said sample in step a), wherein said control tracer comprises a detectable particulate substance having immobilized on its surface a control antigen which binds to said anti-control-antigen antibody.

11. The process of claim 10, wherein said assay tracer and said control tracer are colored particles.

12. The process of claim 11, wherein said colored particles are colored liposomes or colored polymeric beads.

13. The process of claim 12, wherein said colored polymeric beads are colored polystyrene beads.

14. The process of claim 10, wherein said solid support comprises nitrocellulose.

15. The process of claim 10, wherein said first member of said binding pair, immobilized on said assay tracer, is biotin.

16. The process of claim 10, wherein said second member of said binding pair, immobilized in said read-out zone, is avidin, streptavidin, or neutravidin.

17. The process of claim 10, wherein:

a) said lateral flow device further comprises a housing which covers the entire lateral flow device, wherein said housing has

i) an opening over said sample addition area,

ii) a first window for viewing said read-out area, and

iii) a second window for viewing a distal end of said device; and

b) said process further comprises contacting said sample/preparation mixture with said sample addition area by pouring said sample/preparation mixture through said opening.

18. The process of claim 17, wherein an end-of-assay tracer is added to said sample/preparation mixture for viewing through said second window.

19. A kit for determining the presence or amount of an analyte in a sample, comprising:

a) a preparation containing

i) anti-analyte antibodies, which bind to analyte or to both analyte and an analyte analog, and

ii) an assay tracer, wherein said assay tracer comprises a detectable particulate substance having immobilized on its surface, analyte or analyte analog and a first member of a binding pair; and

b) a lateral flow device, comprising a solid support containing, in sequence,

i) said sample addition area,

ii) a capture area, wherein analyte or an analyte analog is immobilized, and

iii) a read-out area comprising at least one zone, wherein a second member of said binding pair, which binds to said first member, is immobilized.

20. The kit of claim 14, wherein said assay tracer is a colored particle.

21. The kit of claim 20, wherein said colored particle is a colored liposome or a colored polymeric bead.

22. The kit of claim 21, wherein said colored polymeric bead is a colored polystyrene bead.

23. The kit of claim 19, wherein said solid support comprises nitrocellulose.

24. The kit of claim 19, wherein said second member of said binding pair, immobilized in said read-out zone, is avidin, streptavidin, or neutravidin.

25. The kit of claim 19, further comprising:

a housing which covers the entire lateral flow device, wherein said housing has

i) an opening over said sample addition area,

ii) a first window for viewing said read-out area, and

iii) a second window for viewing a distal end of said device.

26. The kit of claim 19, further comprising:

a) a second zone in said read-out area, which functions as a control, wherein an anti-control-antigen antibody is immobilized; and

b) a control tracer comprising a detectable particulate substance having immobilized on its surface a control antigen which binds to said anti-control-antigen antibody.

27. The kit of claim 26, wherein said assay tracer and said control tracer are colored particles.

28. The kit of claim 27, wherein said colored particles are colored liposomes or colored polymeric beads.

29. The kit of claim 28, wherein said colored polymeric beads are colored polystyrene beads.

30. The kit of claim 26, wherein said solid support comprises nitrocellulose.

31. The kit of claim 26, wherein said second member of said binding pair, immobilized in said read-out zone, is avidin, streptavidin, or neutravidin.

32. The kit of claim 26, further comprising:

a housing which covers the entire lateral flow device, wherein said housing has

i) an opening over said sample addition area,

ii) a first window for viewing said read-out area, and

iii) a second window for viewing a distal end of said device.