US4293310A
Expired / Lapsed
US
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Photoelectrochemical immunoassay
Assignee: University of Pittsburgh
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Filing: 1980-03-14
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Granted: 1981-10-06
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Expires: 2000-03-14
Abstract
Abstract A method and apparatus for the determination of trace amounts of chemicals in a system, as for example the immunoassay determination of hormones, peptides and drugs in biological fluids, applies photoelectrochemistry to the field of immunoassay. The apparatus and method comprise a photoelectrochemically active molecule as a label in an immunoassay system of labeled analyte, antibody sensitive to analyte, quencher and an electrochemical flow cell with light means. The photoelectrochemically active molecule upon photoexcitation transfers an electron to a quencher molecule; the oxidized molecule is subsequently reduced with an electron from an electrode of the flow cell which is held at suitable potential. This electron is measured as photocurrent. The electrochemical flow cell is channeled for the flow of solution containing photoelectrochemically active species and quencher; and has at least one wall transparent to light such as a laser which is trained on the appropriate region of the cell; and further comprises means to isolate the spectral region of interest and means to distinguish photocurrent from nonphotocurrent. The amount of free labeled analyte in the system is determined by the photocurrent signal.
Inventors
- 👤 Stephen G. Weber
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 12:01
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/533; G01N33/543; G01N33/54366; G01N33/54373; G01N33/5438; G01N33/58; Y; Y10; Y10S; Y10S436/00; Y10S436/805; Y10S436/807; Y10S436/815; Y10S436/816; Y10S436/817
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
117 words
Immunoassay apparatus comprising: (i) a catalytic photoelectrochemical label capable of photoexcitation such that a photocurrent is generated at an electrode of appropriate potential; (ii) a quencher; and (iii) an electrochemical flow cell having, at least one wall transparent to light of a spectrum which will excite said photoelectrochemically active label, and (iv) light means which further comprise, a light source having a spectrum capable of exciting said photoelectrochemically active label, and means for isolating said spectrum, wherein the relative planes of orientation of the electrodes of said cell, and said spectrum are disposed such that said photoelectrochemically active label will react with said quencher to generate said photocurrent, and means for distinguishing between said photocurrent and non-photocurrent.
Claim 2
Dependent Claim
117 words
Immunoassay apparatus comprising: (i) a catalytic photoelectrochemical label capable of photoexcitation such that a photocurrent is generated at an electrode of appropriate potential; (ii) a quencher; and (iii) an electrochemical flow cell having, at least one wall transparent to light of a spectrum which will excite said photoelectrochemically active label, and (iv) light means which further comprise, a light source having a spectrum capable of exciting said photoelectrochemically active label, and means for isolating said spectrum, wherein the relative planes of orientation of the electrodes of said cell, and said spectrum are disposed such that said photoelectrochemically active label will react with said quencher to generate said photocurrent, and means for distinguishing between said photocurrent and non-photocurrent.
Claim 3
Dependent Claim
15 words
The detecting apparatus of claim 1 wherein said photoelectrochemically active label is a ruthenium complex.
Claim 4
Dependent Claim
25 words
The detecting apparatus of claim 2 wherein said ruthenium complex is RuL 3 and L is a derivative of o-phenanthroline or 2,2'-bipyridine or combination thereof.
Claim 5
Dependent Claim
15 words
The detecting apparatus of claim 3 wherein said RuL 3 is tris (2,2'-bipyridyl) ruthenium (ii).
Claim 6
Dependent Claim
15 words
The detecting apparatus of claim 1 wherein said photoelectrochemically active label is an osmium complex.
Claim 7
Dependent Claim
26 words
The detecting apparatus of claim 5 wherein said osmium complex is OsL 3 and L 3 is a derivative of o-phenanthroline or 2,2'-bipyridine or combinations thereof.
Claim 8
Dependent Claim
18 words
The detecting apparatus of claim 1 wherein said quencher is Co(C 2 O 4 ) 3 3- .
Claim 9
Dependent Claim
11 words
The detecting apparatus of claim 1 wherein said quencher is hydrazine.
Claim 10
Dependent Claim
15 words
The detecting apparatus of claim 1 wherein said light source is a hollow cathode lamp.
Claim 11
Dependent Claim
13 words
The detecting apparatus of claim 9 wherein said cathode is a strontium lamp.
Claim 12
Dependent Claim
15 words
The detecting apparatus of claim 1 wherein said light source is a Xe arc lamp.
Claim 13
Dependent Claim
13 words
The detecting apparatus of claim 1 wherein said light source is a laser.
Claim 14
Dependent Claim
27 words
The detecting apparatus of claim 1 wherein said means for distinguishing between said photocurrent and non-photocurrent further comprises a light beam chopper, filters, lenses and lock-in amplifier.
Claim 15
Dependent Claim
50 words
The detecting apparatus of claim 1 wherein said means for distinguishing between said photocurrent and non-photocurrent further comprises a first working electrode exposed to said light and a second working electrode in the dark, such that the difference in current signals is the signal due to the presence of light.
Claim 16
Dependent Claim
17 words
The detecting apparatus of claim 1 wherein said isolated spectrum is in one range at 430-500 nm.
Claim 17
Dependent Claim
17 words
The detecting apparatus of claim 14 wherein said means for isolating said spectrum further comprises a monochromater.
Claim 18
Dependent Claim
23 words
The detecting apparatus of claim 1 wherein said relative planes of orientation are disposed such that said spectrum impinges perpendicularly to said electrodes.
Claim 19
Dependent Claim
22 words
The detecting apparatus of claim 1 wherein said relative planes of orientation are such that said spectrum impinges parallel to said electrodes.
Claim 20
Dependent Claim
109 words
A catalytic photoelectrochemical method of immunoassay comprising the steps of: (i) utilizing a photoelectrochemically labeled antigen and quencher in an assay of analyte, labeled analyte and antibody specific to said analyte, and (ii) measuring the concentration of said photoelectrochemically labeled analyte with an electrochemical flow cell, having means for distinguishing photocurrent and non-photocurrent; light means which further comprises a light source, having a spectrum capable of exciting said photoelectrochemically active label, and means for isolating said spectrum, wherein the relative planes of orientation of the electrodes of said cell, and said spectrum are disposed such that said photoelectrochemically active label will react with said quencher to generate said photocurrent.
Claim 21
Dependent Claim
109 words
A catalytic photoelectrochemical method of immunoassay comprising the steps of: (i) utilizing a photoelectrochemically labeled antigen and quencher in an assay of analyte, labeled analyte and antibody specific to said analyte, and (ii) measuring the concentration of said photoelectrochemically labeled analyte with an electrochemical flow cell, having means for distinguishing photocurrent and non-photocurrent; light means which further comprises a light source, having a spectrum capable of exciting said photoelectrochemically active label, and means for isolating said spectrum, wherein the relative planes of orientation of the electrodes of said cell, and said spectrum are disposed such that said photoelectrochemically active label will react with said quencher to generate said photocurrent.
Claim 22
Dependent Claim
136 words
A catalytic photochemical method of immunoassay comprising the steps of: (i) utilizing a photoelectrochemical as a label with a labeled analyte, and a quencher, in an assay mixture of analyte, labeled analyte and antibodies specific to said analyte, and (ii) measuring the concentration of said photoelectrochemically labeled analyte with an electrochemical flow cell, having, at least one wall transparent to light of a spectrum which will excite said photoelectrochemically active label, means for distinguishing between said photocurrent and nonphotocurrent and light means which further comprises, a light source having a spectrum capable of exciting said photoelectrochemically active label, and means for isolating said spectrum, wherein the relative planes of orientation of the electrodes of said cell, as a spectrum are disposed such that said photoelectrochemically active label will react with said quencher to generate said photocurrent.
Claim 23
Dependent Claim
136 words
A catalytic photochemical method of immunoassay comprising the steps of: (i) utilizing a photoelectrochemical as a label with a labeled analyte, and a quencher, in an assay mixture of analyte, labeled analyte and antibodies specific to said analyte, and (ii) measuring the concentration of said photoelectrochemically labeled analyte with an electrochemical flow cell, having, at least one wall transparent to light of a spectrum which will excite said photoelectrochemically active label, means for distinguishing between said photocurrent and nonphotocurrent and light means which further comprises, a light source having a spectrum capable of exciting said photoelectrochemically active label, and means for isolating said spectrum, wherein the relative planes of orientation of the electrodes of said cell, as a spectrum are disposed such that said photoelectrochemically active label will react with said quencher to generate said photocurrent.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-10-05 | 1985-04-06 | 1985-10-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-10-05 | 1989-04-06 | 1989-10-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-10-05 | 1993-04-06 | 1993-10-06 | $7700.00 | Window Closed / Lapsed |