US4945045A
Expired / Lapsed
US
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Electrochemical methods of assay
Assignee: LABORATOIRES SERONO SA A CORP OF SWITZERLAND
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Filing: 1988-06-23
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Granted: 1990-07-31
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Expires: 2008-06-23
Abstract
Abstract A ligand in a sample can be assayed using electrochemical apparatus containing a working electrode and components comprising: (a) the sample, (b) a specific binding partner to the ligand, or a specific binding partner to the ligand and selected from ligand analogues and specific binding partners, and (c) an electron-source or electron-acceptor; at least one of components (b) being labelled with an electron-transfer mediator capable of aiding the transfer of electrons between the electron-source or electron-acceptor and the working electrode by accepting electrons from the electron-source and donating them to the working electrode, or accepting the electrons from the electrode and donating them to the electron-acceptor. The method includes the step of determining whether, and, if desired, the extent to which, the said transfer of electrons is perturbed by at least one of (i) formation of a complex of said ligand with a specific binding partner and (ii) controlled external influence which produces a perturbation of said transfer of electrons as a function of said complex formation.
Inventors
- 👤 Gordon C. Forrest
- 👤 Simon J. Rattle
- 👤 Grenville A. Robinson
- 👤 Hugh A. O. Hill
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 13:02
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54366; G01N33/54373; G01N33/5438; G01N27/00; G01N27/26; G01N27/28; G01N27/30; G01N27/327; G01N27/3271; Y; Y10; Y10S; Y10S435/00; Y10S435/817; Y10S435/975; Y10S436/00; Y10S436/806
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
164 words
In a homogeneous method of assaying a ligand in a sample using electrochemical apparatus containing a working electrode and components comprising: (a) the sample, and (b) a specific binding partner to the ligand, or a specific binding partner to the ligand and at least one further reagent selected from ligand analogues and specific binding partners to the ligand, the improvement which comprises said components including (c) an electron-source or electron-acceptor selected from oxidoreductase enzymes in cooperation with a substrate therefor and non-enzyme electron sources; one of components (b) being labelled with an electron-transfer mediator capable of aiding the transfer of electrons between the electron-source or electron-acceptor and the working electrode by accepting electrons from the electron-source and donating them to the working electrode, or accepting electrons from said electrode and donating them to the electron-acceptor; and which method includes the step of determining whether the said transfer of electrons is perturbed by formation of a complex of said ligand with a specific binding partner.
Claim 2
Dependent Claim
164 words
In a homogeneous method of assaying a ligand in a sample using electrochemical apparatus containing a working electrode and components comprising: (a) the sample, and (b) a specific binding partner to the ligand, or a specific binding partner to the ligand and at least one further reagent selected from ligand analogues and specific binding partners to the ligand, the improvement which comprises said components including (c) an electron-source or electron-acceptor selected from oxidoreductase enzymes in cooperation with a substrate therefor and non-enzyme electron sources; one of components (b) being labelled with an electron-transfer mediator capable of aiding the transfer of electrons between the electron-source or electron-acceptor and the working electrode by accepting electrons from the electron-source and donating them to the working electrode, or accepting electrons from said electrode and donating them to the electron-acceptor; and which method includes the step of determining whether the said transfer of electrons is perturbed by formation of a complex of said ligand with a specific binding partner.
Claim 3
Dependent Claim
20 words
A method as claimed in claim 1 wherein component (c) is an oxidoreductase enzyme in co-operation with a substrate therefor.
Claim 4
Dependent Claim
17 words
A method as claimed in claim 1 wherein the electron-transfer media comprises ferrocene or a derivative thereof.
Claim 5
Dependent Claim
77 words
A method as claimed in claim 3 wherein the electron-transfer mediator comprises a derivative of ferrocene containing one or more side chains of the formula --CHO, --(CH 2 ) n COOH or --(CH 2 ) m NR 1 R 2 (where n and m are each from 0 to 6) and R 1 and R 2 , which may be the same or different, each represents hydrogen or an alkyl group containing 1 to 4 carbon atoms).
Claim 6
Dependent Claim
35 words
A method as claimed in claim 1 wherein the perturbation in the transfer of electrons is determined from a perturbation in the peak current observed under the application of a preselected potential across the components.
Claim 7
Dependent Claim
16 words
A method as claimed in claim 1 wherein the ligand is an antigen or an antibody.
Claim 8
Dependent Claim
36 words
A method as claimed in claim 1, wherein the extent to which the said transfer of electrons is perturbed is determined, the extent of perturbation being an indication of the amount of ligand in the sample.
Claim 9
Dependent Claim
36 words
A method as claimed in claim 2, wherein the extent to which the said transfer of electrons is perturbed is determined, the extent of perturbation being an indication of the amount of ligand in the sample.
Claim 10
Dependent Claim
15 words
A method as claimed in claim 7, wherein said electron source is ascorbate or NADH.
Claim 11
Dependent Claim
15 words
A method as claimed in claim 1, wherein said electron source is ascorbate or NADH.
Claim 12
Dependent Claim
81 words
A kit for carrying out a method of assay as claimed in claim 1 comprising in at least one container: (i) at least one specific binding partner to the ligand or a specific binding partner to the ligand and at least one further reagent selected from ligand analogues and specific binding partners to the ligand one of the components (i) being labelled with an electron transfer mediator, and (ii) a non-enzyme electron-source capable of donating electrons to said electron transfer mediator.
Claim 13
Dependent Claim
17 words
A kit as claimed in claim 11 which further comprises an electrochemical apparatus containing a working electrode.
Claim 14
Dependent Claim
95 words
A kit for carrying out a method of assay as claimed in claim 2 comprising in separate containers: (i) at least one specific binding partner to the ligand or a specific binding partner to the ligand and at least one further reagent selected from ligand analogues and specific binding partners to the ligand, one of the components (i) being labelled with an electron transfer mediator; and (ii) an oxidoreductase enzyme which in cooperation with a substrate therefor is capable of donating electrons to said electron transfer mediator or accepting electrons from said electron transfer mediator.
Claim 15
Dependent Claim
17 words
A kit as claimed in claim 13 which further comprises an electrochemical apparatus containing a working electrode.
Claim 16
Dependent Claim
25 words
A method as claimed in claim 1 wherein a specific binding partner to the ligand or a ligand analogue is immobilized on the working electrode.
Claim 17
Dependent Claim
106 words
In a heterogeneous method of assaying a ligand in a sample using electrochemical apparatus containing a working electrode and components comprising: (a) the sample, and (b) the specific binding partner to the ligand, or a specific binding partner to the ligand and at least one further reagent selected from ligand analogues and specific binding partners to the ligand, the improvement which comprises said components including (c) an electron-source or electron-acceptor selected from oxidoreductase enzymes in cooperation with a substrate therefor and non-enzyme electron sources; one of components (b) being labelled with an electron-transfer mediator capable of aiding the transfer of electrons between the electron-source or electron-acceptor
Claim 18
Dependent Claim
106 words
In a heterogeneous method of assaying a ligand in a sample using electrochemical apparatus containing a working electrode and components comprising: (a) the sample, and (b) the specific binding partner to the ligand, or a specific binding partner to the ligand and at least one further reagent selected from ligand analogues and specific binding partners to the ligand, the improvement which comprises said components including (c) an electron-source or electron-acceptor selected from oxidoreductase enzymes in cooperation with a substrate therefor and non-enzyme electron sources; one of components (b) being labelled with an electron-transfer mediator capable of aiding the transfer of electrons between the electron-source or electron-acceptor
Claim 19
Dependent Claim
20 words
A method as claimed in claim 16 wherein component (c) is an oxidoreductase enzyme in cooperation with a substrate therefor.
Claim 20
Dependent Claim
17 words
A method as claimed in claim 16 wherein the electron-transfer mediator comprises ferrocene or a derivative thereof.
Claim 21
Dependent Claim
16 words
A method as claimed in claim 16 wherein the ligand is an antigen or an antibody.
Claim 22
Dependent Claim
36 words
A method as claimed in claim 16, wherein the extent to which the said transfer of electrons is perturbed is determined, the extent of perturbation being an indication of the amount of ligand in the sample.
Claim 23
Dependent Claim
24 words
A method as claimed in claim 16 wherein an unlabelled specific binding partner to the ligand under assay is immobilized on a solid support.
Claim 24
Dependent Claim
79 words
A kit for carrying out a method of assay as claimed in claim 16 comprising in at least one container: (i) at least one specific binding partner to the ligand or a specific partner to the ligand and at least one further reagent selected from ligand analogs and specific binding partners to the ligand, one of the components (i) being labelled with an electron-transfer mediator, and (ii) a non-enzyme electron-source capable of donating electrons to said electron transfer mediator.
Claim 25
Dependent Claim
188 words
A kit for carrying out a method of assay as claimed in claim 16 comprising in separate containers: (i) at least one specific binding partner to the ligand or a specific binding partner to the ligand and at least one further reagent selected from ligand analogs and specific binding partners to the ligand, one of the components (i) being labelled with an electron-transfer mediator; and (ii) an oxidoreductase enzyme which in cooperation with a substrate therefor is capable of donating electrons to said electron transfer mediator or accepting electrons from said electron transfer mediator. and the working electrode by accepting electrons from the electron-source and donating them to the working electrode, or accepting electrons from said electrode and donating them to the electron-acceptor; and which method includes the steps of (i) providing means for displacement of any mediator-labelled complex containing said ligand relative to any other mediator-labelled component, or where a mediator-labelled ligand analogue is present as component (b), providing means for displacement of any complex containing said ligand analog relative to non-complexed mediator-labelled ligand analog and (ii) determining whether the said transfer of electrons is thereby perturbed.
References Cited (Backward Citations)
US3966556A
Cited
US4081334A
Cited
US4600690A
Cited
US4340448A
Cited
US4233144A
Cited
US4391904A
Cited
US4319980A
Cited
US4545382A
Cited
EP0078636A1
Cited
EP0125136A2
Cited
EP0125867A2
Cited
EP0125139A2
Cited
US4758323A
Cited
EP0142301A2
Cited
EP0149339A2
Cited
EP0167248A2
Cited
GB8417301D0
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-07-30 | 1994-01-29 | 1994-07-31 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-07-30 | 1998-01-29 | 1998-07-31 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-07-30 | 2002-01-29 | 2002-07-31 | $7700.00 | Window Closed / Lapsed |