Particle bound binding component immunoassay

• US 5,501,949 A
• Filed: 08/15/1994
• Issued: 03/26/1996
• Est. Priority Date: 12/10/1985
• Status: Expired due to Term

• Current Assignee: Murex Diagnostics Corporation (Abbott Laboratories)
• Original Assignee: Murex Diagnostics Corporation (Abbott Laboratories)
• Inventors: Marshall, David L.
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/290,487
• Time in Patent Office: 589 Days
• Field of Search: 422/56-57, 422/101, 427/2, 427/2.11, 435/5, 435/6, 435/7.1, 435/7.2, 435/7.32, 435/7.5, 435/7.92-7.95, 435/970, 435/975, 436/518, 436/523, 436/529-531, 436/532-534, 436/538, 436/810, 436/815, 436/818, 436/824
• US Class Current: 435/5
• CPC Class Codes: G01N 33/538 by sorbent column, particles or resin strip , i.e. sorbent materials
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/56927 Chlamydia
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/815 Test for named compound or class of compounds
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

An immunoassay method for the detection or quantitation of an analyte suspected of being in a solution comprising: (a) combining said specimen, a first binding component, insoluble particles, and second binding component labelled with a signal generating material in a solid phase retention and separation apparatus having a sufficient pore size such that said particles are trapped within said filter yet permitting rapid passage of fluid therethrough in such a manner that an immunological reaction occurs if analyte is present in said specimen, resulting in the formation of an immunocomplex of insolublized first binding component:analyte:second labelled binding component on or within said filter means; (b) separating bound from unbound material; and (c) determining the presence and/or amount of signal produced which is correlative with the amount of analyte present in the solution.

Claims

1. A method for detection or quantitation of an analyte in a solution comprising the steps of:

(a) contacting the solution with insoluble particles having attached thereto a binding component specific for the analyte, so as to form a suspension comprising a first complex wherein the first complex comprises the analyte, the binding component and the insoluble particle;

(b) applying the suspension to at least a portion of a semi-permeable membrane having interstices of dimensions relative to the insoluble particles and having a thickness such that the insoluble particles are retained throughout the thickness of the semi-permeable membrane, the portion of the semi-permeable membrane retaining the insoluble particles defining an assay zone;

(c) contacting the semi-permeable membrane containing the assay zone with a labeling component capable of specifically binding to the first complex so as to form a second complex wherein the second complex comprises the first complex and the labeling component, the semi-permeable membrane permitting the labeling component which is not bound to the first complex to pass out of the assay zone; and

(d) measuring the signal produced by the labeling component of the second complex as an indicator of the presence or amount of the analyte present in the solution.

2. The method of claim 1, further comprising the step of applying to the semi-permeable membrane a wash fluid capable of passing through the semi-permeable membrane after Step (c).

3. The method of claim 1, wherein the semi-permeable membrane comprises a filter.

4. The method of claim 1, wherein the semi-permeable membrane comprises a material selected from the group consisting of glass, synthetic polymers and cellulose.

5. The method of claim 1, wherein the insoluble particles comprise an inert solid phase.

6. The method of claim 5, wherein the inert solid phase comprises a material selected form the group consisting of glass, cellulose, synthetic polymer, and protein.

7. The method of claim 5, wherein the binding component is covalently bound to the solid phase in an amount sufficient to substantially coat the solid phase.

8. The method of claim 5, wherein the binding component is bound to avidin and the solid phase is bound to biotin.

9. The method of claim 1, wherein the analyte is selected from the group consisting of antigens, antibodies, proteins, bacteria, fungi, haptens, hormones, parasites, viruses, and tumor cell markers.

10. The method of claim 1, wherein the analyte is an antigen.

11. The method of claim 10, wherein the antigen is selected from the group consisting of rubella virus, HTLV, cytomegalovirus, herpes virus, Chlamydia, Neisseria gonorrhea and human chorionic gonadotropin.

12. The method of claim 1, wherein the labeling component is selected form the group consisting of enzyme, bioluminescent material, chemiluminescent material, ferromagnetic material and radioactive material.

13. The method of claim 1, wherein the labeling component is alkaline phosphatase.

14. A method for detection or quantitation of an analyte in a solution comprising the steps of:

(a) contacting the solution with insoluble particles having attached thereto a binding component specific for the analyte so as to form a first complex wherein the first complex comprises the analyte, the binding component and the insoluble particle and with a labeling component capable of specifically binding to the first complex so as to form a suspension comprising a second complex, wherein the second complex comprises the first complex and the labeling component;

(b) applying the suspension to at least a portion of a semi-permeable membrane having interstices of dimensions relative to the insoluble particles and having a thickness such that the insoluble particles are retained throughout the thickness of the semi-permeable membrane, the semi-permeable membrane permitting the soluble labeling component which is not bound to the insoluble particles to pass therethrough; and

(c) measuring the signal produced by the labeling component of the second complex as an indicator of the presence or amount of the analyte present in the specimen solution.

15. The method of claim 14, further comprising the step of applying to the semi-permeable membrane a wash fluid capable of passing through the semi-permeable membrane after Step (b).

16. The method of claim 14, wherein the semi-permeable membrane comprises a filter.

17. The method of claim 14, wherein the semi-permeable membrane comprises a material selected from the group consisting of glass, synthetic polymers and cellulose.

18. The method of claim 14, wherein the insoluble particles comprise an inert solid phase.

19. The method of claim 18, wherein the inert solid phase comprises a material selected form the group consisting of glass, cellulose, synthetic polymer, and protein.

20. The method of claim 18, wherein the binding component is covalently bound to the solid phase in an amount sufficient to substantially coat the solid phase.

21. The method of claim 18, wherein the binding component is bound to avidin and the solid phase is bound to biotin.

22. The method of claim 14, wherein the analyte is selected from the group consisting of antigens, antibodies, proteins, bacteria, fungi, haptens, hormones, parasites, viruses, and tumor cell markers.

23. The method of claim 14, wherein the analyte is an antigen.

24. The method of claim 23, wherein the antigen is selected from the group consisting of rubella virus, HTLV, cytomegalovirus, herpes virus, Chlamydia, Neisseria gonorrhea, and human chorionic gonadotropin.

25. The method of claim 14, wherein the labeling component is selected form the group consisting of enzymes, bioluminescent material, chemiluminescent material, ferromagnetic material and radioactive material.

26. The method of claim 14, wherein the labeling component is alkaline phosphatase.