Method and test strip for detecting residues

• US 9,234,889 B1
• Filed: 12/17/2009
• Issued: 01/12/2016
• Est. Priority Date: 12/18/2008
• Status: Active Grant

• Current Assignee: Charm Sciences, Inc.
• Original Assignee: Charm Sciences, Inc.
• Inventors: Saul, Steven J., Douglas, David W.
• Primary Examiner(s): Crawford, Erik B
• Application Number:
  US12/640,764
• Time in Patent Office: 2,217 Days
• Field of Search: 436/514, 436/518, 436524-534, 436/536, 436538-541, 436 20- 23, 436/924, 435/7.1, 4352871-2873, 4352877-2879, 435/970
• US Class Current: 1/1
• CPC Class Codes: G01N 2333/38 from Aspergillus
• CPC Class Codes: G01N 2333/385 from Penicillium
• CPC Class Codes: G01N 33/537 with separation of immune complex from unbound antigen or antibody
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• 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 33/9446 Antibacterials

Abstract

A method and test strip for detecting one or more analytes in a test solution. The method and test strip include a detectable component, such as a gold particle, that is linked to a binder, such as an antibody, through a linkage. The linkage can include an antibody binding protein and an antispecies antibody.

Claims

1. A lateral flow test strip for detecting an analyte in a test solution, the test strip comprising:

(a) a detectable component, the detectable component linked to a binder through a linkage to form a detectable binder, the detectable binder configured for generating a detectable signal, the linkage comprising;

(i) an antibody binding protein; and

(ii) an anti-species antibody, the anti-species antibody having affinity to the binder, and the antibody binding protein having affinity to both the detectable component and the anti-species antibody, and wherein the detectable binder is further configured to combine with the analyte from the solution to, through affinity of the binder, form a detectable binder-analyte complex;

(b) a test zone on the test strip, the test zone comprising a first capture agent immobilized thereon, whereby said first capture agent captures the detectable binder when the detectable binder has not formed the detectable binder-analyte complex;

(c) a control zone on the test strip, the control zone comprising a second capture agent immobilized thereon, wherein the second capture agent comprises an antibody to the anti-species antibody, and whereby said second capture agent can capture the detectable binder whether or not the detectable binder formed the detectable binder-analyte complex; and

wherein the test strip is configured to allow the test solution and the detectable binder to flow first to the test zone and second to the control zone, and wherein capturing of the detectable binder at either the test zone or control zone generates a detectable signal and whereby a greater signal in the control zone, as compared to the test zone, indicates a positive result.

2. The lateral flow test strip of claim 1 wherein the antibody binding protein comprises protein A.

3. The lateral flow test strip of claim 1, wherein the analyte binder comprises an antibody.

4. The lateral flow test strip of claim 1 wherein the analyte comprises ochratoxin and wherein the analyte binder comprises an antibody to ochratoxin.

5. The lateral flow test strip of claim 1 wherein the first capture agent comprises a representative analyte.

6. The lateral flow test strip of claim 1, wherein the first capture agent is immobilized on a solid support through a carrier protein.

7. The lateral flow test strip of claim 6 wherein the carrier protein comprises bovine serum albumin.

8. The lateral flow test strip of claim 1, wherein the analyte comprises cloxacillin and wherein the analyte binder comprises an antibody to cloxacillin.

9. The lateral flow test strip of claim 1, wherein the analyte binder comprises a monoclonal antibody.

10. The lateral flow test strip of claim 1, wherein the analyte binder comprises a polyclonal antibody.

11. The lateral flow test strip of claim 1 wherein the anti-species antibody comprises a polyclonal antibody.

12. The lateral flow test strip of claim 1 wherein the anti-species antibody comprises a rabbit antibody.

13. The lateral flow test strip of claim 1 wherein the anti-species antibody comprises a rabbit anti-mouse antibody.

14. The lateral flow test strip of claim 1 wherein the anti-species antibody comprises a rabbit anti-sheep antibody.

15. The lateral flow test strip of claim 1 wherein the second capture agent comprises a goat anti-rabbit antibody.

16. The lateral flow test strip of claim 1 wherein the detectable component comprises colloidal gold.

17. The lateral flow test strip of claim 1, wherein said test strip is configured such that, on application, the test solution solubilizes the detectable binder and migrates through the lateral flow test strip by the forces of lateral capillary flow.

18. A lateral-flow test method for detecting one or more analytes in a test sample, the method comprising:

(a) adding the test sample to the lateral flow test strip of claim 1;

(b) allowing the sample to contact the detectable binder, so that the detectable binder reacts with the analyte to form the detectable binder-analyte complex;

(c) allowing the detectable binder to come in contact with the first capture agent, wherein the detectable binder will react with the first capture agent generating the detectable signal, and wherein the test is negative for the detection of the one or more analytes when the signal from the test zone is greater than the signal in the control zone and the test is positive for the one or more analytes when the signal from the control zone is greater than the signal from the test zone.

19. The method of claim 18 wherein the antibody binding protein comprises Protein A.

20. The method of claim 18, wherein the binder comprises an antibody.

21. The method of claim 20 wherein the analyte comprises ochratoxin and wherein the antibody comprises an antibody to ochratoxin.

22. The lateral flow test strip of claim 18, wherein the detectable binder is applied to the test solution prior to being applied to the test strip.

23. The lateral flow test strip of claim 18, wherein the analyte comprises cloxacillin and wherein the analyte binder comprises an antibody to cloxacillin.

24. The lateral flow test strip of claim 18, wherein the analyte binder comprises a monoclonal antibody.

25. The lateral flow test strip of claim 18, wherein the analyte binder comprises a polyclonal antibody.

26. The lateral flow test strip of claim 18, wherein the anti-species antibody comprises a polyclonal antibody.

27. The lateral flow test strip of claim 18, wherein the anti-species antibody comprises a rabbit antibody.

28. The lateral flow test strip of claim 18, wherein the anti-species antibody comprises a rabbit anti-mouse antibody.

29. The lateral flow test strip of claim 18, wherein the anti-species antibody comprises a rabbit anti-sheep antibody.

30. A lateral flow test strip for detecting an analyte in a test solution, the test strip comprising:

(a) a detectable component, the detectable component linked to a binder through a linkage to form a detectable binder, the detectable binder configured for generating a detectable signal, the linkage comprising;

(i) an antibody binding protein; and

(ii) an anti-species antibody, the anti-species antibody having affinity to the binder, and the antibody binding protein having affinity to both the detectable component and the anti-species antibody, and wherein the detectable binder is further configured to combine with the analyte from the solution to, through affinity of the binder, form a detectable binder-analyte complex;

(b) a test zone on the test strip, the test zone comprising a first capture agent immobilized thereon, whereby said first capture agent captures the detectable binder when the detectable binder has not formed the detectable binder-analyte complex;

(c) a control zone on the test strip, the control zone comprising a second capture agent immobilized thereon, wherein the second capture agent comprises an anti-rabbit antibody with affinity to the anti-species antibody, and whereby said second capture agent can capture the detectable binder whether or not the detectable binder formed the detectable binder-analyte complex; and

wherein the test strip is configured to allow the test solution and the detectable binder to flow first to the test zone and second to the control zone, and wherein capturing of the detectable binder at either the test zone or control zone generates a detectable signal and whereby a greater signal in the control zone, as compared to the test zone, indicates a positive result.