Method and apparatus for conducting electrochemiluminescence measurements

• US 5,147,806 A
• Filed: 09/27/1991
• Issued: 09/15/1992
• Est. Priority Date: 04/29/1988
• Status: Expired due to Term

• Current Assignee: BioVeris Corporation (Roche Holding AG)
• Original Assignee: IGEN, Inc. (Teligent Incorporated)
• Inventors: Kamin, Ralph A., Leland, Jonathan K., Hall, Lee O.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): ALEXANDER, LYLE
• Application Number:
  US07/773,971
• Time in Patent Office: 354 Days
• Field of Search: 422/52, 422/81, 435/808, 436/172, 436/149
• US Class Current: 436/149
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 27/305 optically transparent or photoresponsive electrodes
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus

Abstract

A method and apparatus for conducting electrochemiluminescence (ECL) measurements which control the initial conditions relating to the surface state of a triggering working electrode by reproducibly creating and maintaining a favorable surface condition so as to enhance the precision and detection limit of the measurements. Assays are performed with ECL detection in a flow-through cell environment and the precision and detection limit are enhanced by alternating initialization and measurement steps.

Claims

1. A method of conducting an assay by measuring the luminescence emitted by en electrochemiluminescent moiety in a solution and being especially adapted for the assay of a biological material comprising the steps of

(a) cleaning and/or conditioning a working electrode by applying an initializing voltage waveform to the working electrode in the presence of one or more initializing solutions to obtain a specific surface condition on said working electrode;

(b) maintaining the specific surface condition of the working electrode by applying a preoperative voltage waveform to the working electrode until the initiation of the measurement step (d); and

(c) measuring the electrochemiluminescence emitted by said electrochemiluminescent moiety by

(i) activating the electrochemiluminescent moiety in solution by applying an electrochemiluminescence-inducing voltage waveform to the working electrode in the presence of the moiety and

(ii) measuring the luminescence emitted by the electrochemiluminescent moiety.

2. A method according to claim 1 wherein the specific surface condition is a redox state selected with respect to said solution that includes the electrochemiluminescent moiety.

3. A method according to claim 1 wherein said initializing comprises passing fluid pulses of a cleaning and/or conditioning solution across said working electrode separated by gas pulse.

4. A method according to claim 1 wherein said initializing solution comprises a cleaning solution.

5. A method according to claim 1 wherein said initializing solution comprises a conditioning solution.

6. A method according to claim 1 wherein said initilizing solution comprises a cleaning and conditioning solution.

7. A method according to claim 1, wherein said measurement step includes a plurality of said electrochemiluminescence-inducing voltage waveforms.

8. A method according to claim 1, wherein said working electrode is continuously exposed to one or more initializing solutions while said initializing voltage waveform is continuously applied to it.

9. A method according to claim 1 wherein the solution that includes the electrochemiluminescent moiety flows during said measuring step.

10. A method according to claim 1, wherein said step of cleaning and/or conditioning includes a step of cleaning said working electrode by applying a first voltage waveform thereto in the presence of a cleaning solution and a step of conditioning said working electrode by applying a second voltage waveform thereto in the presence of a conditioning solution.

11. A method according to claim 10 wherein said cleaning solution is the same as said conditioning solution.

12. A method according to claim 10 wherein said cleaning solution is different from said conditioning solution.

13. A method according to claim 1 wherein said specific surface condition comprises a redox state selected with respect to the material of said working electrode.

14. A method according to claim 13, wherein said redox state is an oxidized state.

15. A method according to claim 14 wherein said working electrode comprises platinum.

16. A method according to claim 13 wherein said redox state is a reduced state.

17. A method according to claim 16 wherein said working electrode comprises gold.

18. A method according to claim 1, wherein the solution that includes the electrochemiluminescent moiety also includes a biological matrix.

19. A method according to claim 18 wherein said solution is flowing during said measurement step.

20. A method according to claim 7 wherein said solution includes a tertiary alkylamine.

21. A method according to claim 20 wherein said tertiary alkylamine comprises tripropylamine.

22. A method according to claim 20 wherein said solution comprises a buffer component.

23. An apparatus for conducting an assay by measuring the luminescence emitted by an electrochemiluminescent moiety and being especially adapted for the assay of a biological material, comprising

(a) cell means for receiving a solution containing an electrochemiluminescent moiety,

(b) working electrode means operatively associated with said cell means and adapted for operative exposure to said solution,

(c) counterelectrode means adapted for operative exposure to said solution,

(d) means for obtaining a specific surface condition on said working electrode means by applying an initializing voltage waveform to said working electrode means comprising a source of an initializing voltage waveform,

(e) means for maintaining said specific surface condition by applying a preoperative voltage waveform to said working electrode means comprising a source of a preoperative voltage waveform and

(f) means for activating electrochemiluminescent moiety in the cell means by applying an electrochemiluminescence-inducing voltage waveform to said working electrode means comprising a source of an electrochemiluminescence-inducing voltage waveform,

said means recited in (d), (e) and (f) comprises voltage control means constructed for operative connection to a voltage source and to said working electrode means, and

(g) means for measuring the luminescence emitted by said electrochemiluminescent moiety.

24. An apparatus according to claim 23 wherein said apparatus further comprises fluid transport means.

25. An apparatus according to claim 23 wherein the specific surface condition is a redox state selected with respect to said solution that includes the electrochemiluminescent moiety.

26. An apparatus according to claim 23 wherein the cell means is constructed for receiving a solution the includes the electrochemiluminescent moiety and a biological matrix.

27. An apparatus according to claim 23 wherein said specific surface condition comprises a redox state selected with respect to the material of said working electrode means.

28. An apparatus according to claim 27 wherein said redox state is an oxidized state.

29. An apparatus according to claim 28 wherein said working electrode comprises platinum.

30. An apparatus according to claim 27 wherein said redox state is a reduced state.

31. An apparatus according to claim 30 wherein said working electrode means comprises gold.