Modular assembly for reagentless affinity separation and detection of analyte

• US 6,660,532 B1
• Filed: 12/16/1999
• Issued: 12/09/2003
• Est. Priority Date: 04/11/1997
• Status: Expired due to Term

• Current Assignee: Agency For Science Technology and Research
• Original Assignee: Science & Technology Corporation
• Inventors: Tender, Leonard, Yan, Juchao, Hampton, Philip, Opperman, Kimberly, Sklar, Larry A., Lopez, Gabriel P., Rabinovich, Emmanuel, Jain, Ravinder K.
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Padmanabhan, Kartic
• Application Number:
  US09/402,682
• Time in Patent Office: 1,454 Days
• Field of Search: 436/501, 436/164, 436/172, 436/518, 436/528, 436/805, 436/149, 436/151, 436/165, 436/525, 436/527, 436/537, 436/806, 422/68.1, 422/82.05, 422/82.08, 422/55, 422/57, 422/82.01, 435/7.1, 435/7.8, 435/7.92, 435/287.1, 435/287.2, 435/288.7, 435/808, 204/193, 204/403
• US Class Current: 436/518
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/553 Metal or metal coated
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property

Abstract

The invention provides a reagentless assay kit for analyte in a sample comprising a modular affinity assembly including at least one sensor unit comprising a ligand having binding affinity for the analyte (affinity module) operatively associated with a reporter probe (reporter module) responsive to changes in the sensor unit induced by analyte/receptor complex formation by transduction of a characteristic detectable signal. Assays employing the modular assembly are also provided.

Claims

1. A modular assembly for affinity separation of an analyte from a sample comprising:

an affinity support comprised of a metal; and

at least one sensor unit comprising;

an affinity module, said affinity module having a binding affinity for said analyte; and

a signal-competent reporter module, said signal competent reporter module being directly or indirectly signal-responsive to complex formation between said affinity module and said analyte, wherein said signal competent reporter molecule is immobilized on said affinity support, and said signal-competent reporter module being spaced from said affinity support such that said complex formation provides a change in the distance between said signal-competent reporter module and said affinity support.

2. The modular assembly of claim 1, wherein the reporter module is indirectly responsive to complex formation and the modular assembly further includes an effector module associated with the affinity support for mediating reporter module signal response to complex formation.

3. The modular assembly of claim 1, wherein the reporter module comprises a luminophor.

4. The modular assembly of claim 2, wherein the reporter module comprises a luminophor.

5. The modular assembly of claim 3, wherein the signal response of the reporter module is directly induced by analyte binding.

6. The modular assembly of claim 5, wherein the signal response is induced by collisional quenching, photoinduced electron transfer, or resonance energy transfer.

7. The modular assembly of claim 4, wherein signal response of the reporter module is induced by displacement of this module relative to the effector module on complex formation.

8. The modular assembly of claim 7, wherein the signal response is induced by a change in the dielectric environment of the reporter module.

9. The modular assembly of claim 7, wherein the effector module comprises a metallic component.

10. The modular assembly of claim 1, wherein the affinity and reporter modules are immobilized on the support in spaced relationship.

11. An assay kit for detection of an analyte comprising the modular assembly of one of claims 1-10.

12. A method for detecting an analyte in a sample comprising exposing the modular assembly of one of claims 1-10 to the sample and detecting a signal from the reporter module.

13. The modular assembly of claim 1, wherein the reporter module comprises an electrophor.

14. The modular assembly of claim 2, wherein the reporter module comprises an electrophor.

15. The modular assembly of claim 3, wherein said change in the distance between said signal-competent reporter module and said support is measured from the luminescent center of said luminophor to said support.

16. The modular assembly of claim 4, wherein said change in the distance between said signal-competent reporter module and said support is measured from the luminescent center of said luminophor to said support.

17. The modular assembly of claim 13, wherein said electrophor is a redox reporter.

18. The modular assembly of claim 17, wherein said redox reporter is viologen.

19. The modular assembly of claim 14, wherein said electrophor is a redox reporter.

20. The modular assembly of claim 19, wherein said redox reporter is viologen.

21. The modular assembly of claim 13, wherein said electrophor is a redox enzyme.

22. The modular assembly of claim 21, wherein said redox enzyme is horseradish peroxide.

23. The modular assembly of claim 14, wherein said electrophor is a redox enzyme.

24. The modular assembly of claim 23, wherein said redox enzyme is horseradish peroxide.

25. The modular assembly of claim 1, wherein said affinity module is affixed to said support.

26. The modular assembly of claim 1, wherein said affinity module is affixed to said support and spaced apart from said signal-competent reporter module.

27. The modular assembly of claim 1, wherein said affinity module is affixed to said support and linked to said signal-competent reporter module.

28. The modular assembly of claim 1, wherein said signal-competent reporter module is spaced from said support such that fluorescence is not enched.