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US5527684A
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Method of measuring the luminescence emitted in a luminescent assay
Assignee: CIS Bio International SA
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Filing: 1992-01-27
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Granted: 1996-06-18
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Expires: 2012-01-27
Abstract
Abstract PCT No. PCT/FR92/00069 Sec. 371 Date Jul. 28, 1993 Sec. 102(e) Date Jul. 28, 1993 PCT Filed Jan. 27, 1992 PCT Pub. No. WO92/13264 PCT Pub. Date Aug. 6, 1992.The present invention relates to a method employing at least one luminescent tracer compound and a luminescent compound used as an internal reference, which, when exposed to the same excitation wavelength, are capable of emitting at difference wavelengths, lambda 2 and lambda 1 respectively, either by direct luminescence or by the induction of a luminescent emission, and correcting the measurement of the luminescence emitted by the tracer compound at wavelength lambda 2 on the basis of the measurement of the luminescence emitted by the reference compound at wavelength lambda 1.
Inventors
- 👤 Michel Mabile
- 👤 Gerard Mathis
- 👤 Etienne J.-P. Jolu
- 👤 Dominique Pouyat
- 👤 Christophe Dumont
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 23:06
CPC Classifications
G; G01; G01N; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/6428; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582; G01N2021/6417; G01N2021/6421; G01N2021/6439; G01N2021/6441; G01N21/17; G01N21/25; G01N21/27; G01N21/274; Y; Y10; Y10S; Y10S435/00; Y10S435/962; Y10S435/968; Y10S435/973; Y10S436/00; Y10S436/825; Y10T; Y10T436/00; Y10T436/13
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
230 words
A method for detecting or determining a target analyte in a sample in which it may be present by measurement of emitted luminescence in a chemiluminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a tracer compound comprising a receptor capable of specifically binding to the target analyte conjugated to a luminescent label capable of generating emitted luminescence at a measuring wavelength λ 2 in the presence of (iii) an internal reference compound capable of emitting luminescence at a correcting wavelength λ 1 , in order to form a specific binding complex between the target analyte and the tracer, wherein the tracer compound and the internal reference compound are the same or different; (b) irradiating the reaction mixture at a single excitation wavelength by an external radiation source; (c) simultaneously measuring both the emitted luminescence at λ 2 due to the tracer bound in the specific binding complex and the emitted luminescence at λ 1 due to the internal reference compound which takes account of interference parameters of the reaction mixture; (d) calculating a corrected luminescence for the luminescence emitted by the tracer compound at λ2 based on the luminescence emitted by the internal reference compound at λ 1 ; and (e) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 2
Independent Claim
230 words
A method for detecting or determining a target analyte in a sample in which it may be present by measurement of emitted luminescence in a chemiluminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a tracer compound comprising a receptor capable of specifically binding to the target analyte conjugated to a luminescent label capable of generating emitted luminescence at a measuring wavelength λ 2 in the presence of (iii) an internal reference compound capable of emitting luminescence at a correcting wavelength λ 1 , in order to form a specific binding complex between the target analyte and the tracer, wherein the tracer compound and the internal reference compound are the same or different; (b) irradiating the reaction mixture at a single excitation wavelength by an external radiation source; (c) simultaneously measuring both the emitted luminescence at λ 2 due to the tracer bound in the specific binding complex and the emitted luminescence at λ 1 due to the internal reference compound which takes account of interference parameters of the reaction mixture; (d) calculating a corrected luminescence for the luminescence emitted by the tracer compound at λ2 based on the luminescence emitted by the internal reference compound at λ 1 ; and (e) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 3
Independent Claim
261 words
A method for detecting or determining a target analyte in a sample in which it may be present by measurement of emitted luminescence in a luminescent assay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a tracer compound comprising a receptor capable of specifically binding to the target analyte conjugated to a luminescent label, said tracer compound capable of generating emitted luminescence at a measuring wavelength λ 2 in the presence of (iii) an internal reference compound capable of generating emitted luminescence at a correcting wavelength λ 1 , in order to form a specific binding complex between the target analyte and the tracer, wherein the emitted luminescences may be generated by direct luminescence or by induction of a luminescent emission by non-radiative energy transfer, and wherein the tracer compound and the internal reference compound are the same or different; (b) simultaneously irradiating the trace compound and the internal reference compound at a single excitation wavelength by an external radiation source; (c) measuring both the emitted luminescence at λ 2 due to the tracer bound in the specific binding complex and the emitted luminescence at λ 1 due to the internal reference compound which takes account of interference parameters of the reaction mixture; (d) calculating a corrected luminescence for the luminescence emitted by the tracer compound at λ2 based on the luminescence emitted by the internal reference compound at λ 1 ; and (e) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 4
Independent Claim
261 words
A method for detecting or determining a target analyte in a sample in which it may be present by measurement of emitted luminescence in a luminescent assay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a tracer compound comprising a receptor capable of specifically binding to the target analyte conjugated to a luminescent label, said tracer compound capable of generating emitted luminescence at a measuring wavelength λ 2 in the presence of (iii) an internal reference compound capable of generating emitted luminescence at a correcting wavelength λ 1 , in order to form a specific binding complex between the target analyte and the tracer, wherein the emitted luminescences may be generated by direct luminescence or by induction of a luminescent emission by non-radiative energy transfer, and wherein the tracer compound and the internal reference compound are the same or different; (b) simultaneously irradiating the trace compound and the internal reference compound at a single excitation wavelength by an external radiation source; (c) measuring both the emitted luminescence at λ 2 due to the tracer bound in the specific binding complex and the emitted luminescence at λ 1 due to the internal reference compound which takes account of interference parameters of the reaction mixture; (d) calculating a corrected luminescence for the luminescence emitted by the tracer compound at λ2 based on the luminescence emitted by the internal reference compound at λ 1 ; and (e) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 5
Dependent Claim
31 words
The method according to claim 2 wherein the luminescence emitted by the internal reference compound and by the tracer compound, at wavelengths λ 1 and λ 2 respectively, are measured simultaneously.
Claim 6
Dependent Claim
31 words
The method according to claim 2 wherein the step of calculating a corrected luminescence includes dividing the luminescence at λ2 by the luminescence at λ 1 to obtain the corrected luminescence.
Claim 7
Dependent Claim
105 words
The method according to claim 2 wherein the luminescence emitted by the internal reference compound at wavelength λ 1 and the luminescence emitted by the tracer compound at wavelength λ 2 are measured by a multi-channel luminescent measuring device, and wherein the correction of the measurement of the luminescence emitted by the tracer compound at wavelength λ 2 includes fixing a counting rate on a channel measuring the luminescent emission of the internal reference compound at wavelength λ 1 , and then, when this rate is reached, triggering the end of the measurement on a channel measuring the luminescent emission at wavelength λ 2 .
Claim 8
Dependent Claim
18 words
The method according to claim 2 wherein the tracer compound and the internal reference compound are the same.
Claim 9
Dependent Claim
17 words
The method according to claim 2 wherein the tracer compound and the internal reference compound are different.
Claim 10
Dependent Claim
18 words
The method according to claim 2 wherein the tracer compound and/or the internal reference compound are fluorescent compounds.
Claim 11
Dependent Claim
21 words
The method according to claim 2 wherein the tracer compound and/or the internal reference compound are rare earth chelates or cryprates.
Claim 12
Dependent Claim
29 words
The method according to claim 2 wherein the emission lifetime of the tracer compound and/or the emission lifetime of the internal reference compound are of more than one microsecond.
Claim 13
Dependent Claim
17 words
The method according to claim 2 wherein the corrected luminescence is measured with a picomol/liter measuring sensitivity.
Claim 14
Independent Claim
275 words
A method for detecting or determining a target analyte in a serum sample in which it may be present by measurement of emitted luminescence in a luminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a tracer compound comprising a receptor capable of specifically binding to the target analyte conjugated to a luminescent label, said tracer compound capable of generating emitted luminescence at a measuring wavelength λ 2 , wherein the emitted luminescence may be generated by direct luminescence or by induction of a luminescent emission by non-radiative energy transfer to the receptor of said analyte conjugated with a luminescent acceptor compound (iii) in the presence of an internal reference compound capable of generating emitted luminescence at a correcting wavelength λ 1 , in order to form a specific binding complex between the target analyte and the tracer, and wherein the tracer compound and the internal reference compound are the same or different; (b) simultaneously irradiating the tracer compound and the internal reference compound at a single excitation wavelength by an external radiation source; (c) measuring both the emitted luminescence at λ 2 due to the tracer bound in the specific binding complex and the emitted luminescence at λ 1 due to the internal reference compound which takes account of interference parameters of the reaction mixture; (d) calculating a corrected luminescence for the luminescence emitted by the tracer compound at λ2 based on the luminescence emitted by the internal reference compound at λ 1 ; and (e) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 15
Independent Claim
275 words
A method for detecting or determining a target analyte in a serum sample in which it may be present by measurement of emitted luminescence in a luminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a tracer compound comprising a receptor capable of specifically binding to the target analyte conjugated to a luminescent label, said tracer compound capable of generating emitted luminescence at a measuring wavelength λ 2 , wherein the emitted luminescence may be generated by direct luminescence or by induction of a luminescent emission by non-radiative energy transfer to the receptor of said analyte conjugated with a luminescent acceptor compound (iii) in the presence of an internal reference compound capable of generating emitted luminescence at a correcting wavelength λ 1 , in order to form a specific binding complex between the target analyte and the tracer, and wherein the tracer compound and the internal reference compound are the same or different; (b) simultaneously irradiating the tracer compound and the internal reference compound at a single excitation wavelength by an external radiation source; (c) measuring both the emitted luminescence at λ 2 due to the tracer bound in the specific binding complex and the emitted luminescence at λ 1 due to the internal reference compound which takes account of interference parameters of the reaction mixture; (d) calculating a corrected luminescence for the luminescence emitted by the tracer compound at λ2 based on the luminescence emitted by the internal reference compound at λ 1 ; and (e) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 16
Dependent Claim
13 words
The method according to claim 12 wherein the method is a homogeneous method.
Claim 17
Dependent Claim
32 words
The luminescent immunoassay according to claim 12 wherein the luminescence emitted by the internal reference compound and by the tracer compound, at wavelength λ 1 and λ 2 respectively, are measured simultaneously.
Claim 18
Dependent Claim
104 words
The method according to claim 12 wherein the tracer compound comprising a receptor capable of specifically binding to the target analyte conjugated to a luminescent acceptor compound and the internal reference compound comprises the analyte conjugated to a luminescent donor compound; step (a) further comprises incubating said reaction mixture after either sequential or simultaneous addition of the tracer compound and the internal reference compound; step (b) occurs at the excitation wavelength of the luminescent donor compound; and step (c) comprises measuring the luminescent emitted by the luminescent donor compound at λ 1 and the luminescence emitted by the accepter compound at λ 2 .
Claim 19
Dependent Claim
246 words
An excess method for detecting or determining a target analyte in a serum sample in which it may be present by measurement of emitted luminescence in a luminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a first reagent comprising a first receptor for the target analyte, coupled with a luminescent donor compound, said first receptor capable of specifically binding to the target analyte and (iii) a second reagent comprising a second receptor capable of specifically binding to the target analyte, said second reagent being coupled with a luminescent acceptor compound, said donor compound capable of transferring energy to said acceptor compound, said donor compound and said acceptor compound capable of generating emitted luminescence at measuring wavelengths λ 1 and λ 2 respectively; (b) incubating said mixture after addition of each reagent or after addition of both reagents; (c) irradiating the resulting mixture at an excitation wavelength of the donor compound; (d) measuring the emitted luminescence of the donor compound at wavelength λ 1 , this measurement serving as a reference, and the emitted luminescence of the acceptor compound at wavelength λ 2 resulting from any energy transfer; (e) calculating a corrected luminescence for the luminescence emitted by the acceptor compound at λ 2 based on the luminescence emitted by the donor compound at λ 1 ; and (f) correlating the corrected luminescence reading the presence or amount of the target analyte in the sample.
Claim 20
Dependent Claim
246 words
An excess method for detecting or determining a target analyte in a serum sample in which it may be present by measurement of emitted luminescence in a luminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a first reagent comprising a first receptor for the target analyte, coupled with a luminescent donor compound, said first receptor capable of specifically binding to the target analyte and (iii) a second reagent comprising a second receptor capable of specifically binding to the target analyte, said second reagent being coupled with a luminescent acceptor compound, said donor compound capable of transferring energy to said acceptor compound, said donor compound and said acceptor compound capable of generating emitted luminescence at measuring wavelengths λ 1 and λ 2 respectively; (b) incubating said mixture after addition of each reagent or after addition of both reagents; (c) irradiating the resulting mixture at an excitation wavelength of the donor compound; (d) measuring the emitted luminescence of the donor compound at wavelength λ 1 , this measurement serving as a reference, and the emitted luminescence of the acceptor compound at wavelength λ 2 resulting from any energy transfer; (e) calculating a corrected luminescence for the luminescence emitted by the acceptor compound at λ 2 based on the luminescence emitted by the donor compound at λ 1 ; and (f) correlating the corrected luminescence reading the presence or amount of the target analyte in the sample.
Claim 21
Dependent Claim
235 words
A competitive method for detecting or determining a target analyte in a serum sample in which it may be present by measurement of emitted luminescence in a luminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a first reagent comprising a first receptor for said target analyte, coupled with a luminescent donor compound, said first receptor capable of specifically binding to the target analyte and (iii) a second reagent comprising the target analyte coupled with a luminescent acceptor compound, said donor compound capable of transferring energy to said acceptor compound, said donor compound and said acceptor compound capable of generating emitted luminescence at measuring wavelength λ 1 and λ 2 respectively; (b) incubating said mixture after addition of each reagent or after addition of both reagents; (c) irradiating the resulting mixture at an excitation wavelength of the donor compound; (d) measuring the emitted luminescence of the donor compound at wavelength λ 1 , this measurement serving as a reference, and the emitted luminescence of the acceptor compound at wavelength λ 2 resulting from any energy transfer; (e) calculating a corrected luminescence for the luminescence emitted by the acceptor compound at λ 2 based on the emitted luminescence by the donor compound at λ 1 ; and (f) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 22
Dependent Claim
235 words
A competitive method for detecting or determining a target analyte in a serum sample in which it may be present by measurement of emitted luminescence in a luminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a first reagent comprising a first receptor for said target analyte, coupled with a luminescent donor compound, said first receptor capable of specifically binding to the target analyte and (iii) a second reagent comprising the target analyte coupled with a luminescent acceptor compound, said donor compound capable of transferring energy to said acceptor compound, said donor compound and said acceptor compound capable of generating emitted luminescence at measuring wavelength λ 1 and λ 2 respectively; (b) incubating said mixture after addition of each reagent or after addition of both reagents; (c) irradiating the resulting mixture at an excitation wavelength of the donor compound; (d) measuring the emitted luminescence of the donor compound at wavelength λ 1 , this measurement serving as a reference, and the emitted luminescence of the acceptor compound at wavelength λ 2 resulting from any energy transfer; (e) calculating a corrected luminescence for the luminescence emitted by the acceptor compound at λ 2 based on the emitted luminescence by the donor compound at λ 1 ; and (f) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 23
Independent Claim
301 words
A method for detecting or determining a target analyte in a serum sample in which it may be present by measurement of emitted luminescence in a luminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a first reagent comprising a first receptor capable of specifically binding to the target analyte, (iii) a second reagent comprising either the target analyte or a second receptor capable of specifically binding to the target analyte, and (iv) a luminescent compound capable of emitting luminescence at wavelength λ 1 as an internal reference compound, wherein one of (ii) or (iii) is coupled to a fluorescent tracer compound capable of generating emitted luminescence at wavelength λ 2 and the other is coupled to a heavy atom or moieties containing a heavy atom such that the heavy atom induces an increase in spin-orbit coupling of the fluorescent tracer compound thereby modulating emitted luminescence generated by the fluorescent tracer compound; (b) incubating the resulting medium either after addition of each reagent or after addition of all reagents; (c) simultaneously irradiating the mixture at a single excitation wavelength common to both the fluorescent tracer compound and the internal reference compound by an external radiation source; (d) measuring the emitted luminescence of the fluorescent tracer compound at wavelength λ 2 , and the emitted luminescence of the internal reference compound at wavelength λ 1 , said emitted luminescence at a wavelength λ 2 being modulated by the heavy atom effect; (e) calculating a corrected luminescence for the luminescence emitted by the fluorescent tracer compound at λ 2 based on the luminescence emitted by the internal reference compound at λ 1 ; and (f) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
Claim 24
Independent Claim
301 words
A method for detecting or determining a target analyte in a serum sample in which it may be present by measurement of emitted luminescence in a luminescent immunoassay, said method comprising the steps of: (a) forming a reaction mixture by contacting (i) the sample with (ii) a first reagent comprising a first receptor capable of specifically binding to the target analyte, (iii) a second reagent comprising either the target analyte or a second receptor capable of specifically binding to the target analyte, and (iv) a luminescent compound capable of emitting luminescence at wavelength λ 1 as an internal reference compound, wherein one of (ii) or (iii) is coupled to a fluorescent tracer compound capable of generating emitted luminescence at wavelength λ 2 and the other is coupled to a heavy atom or moieties containing a heavy atom such that the heavy atom induces an increase in spin-orbit coupling of the fluorescent tracer compound thereby modulating emitted luminescence generated by the fluorescent tracer compound; (b) incubating the resulting medium either after addition of each reagent or after addition of all reagents; (c) simultaneously irradiating the mixture at a single excitation wavelength common to both the fluorescent tracer compound and the internal reference compound by an external radiation source; (d) measuring the emitted luminescence of the fluorescent tracer compound at wavelength λ 2 , and the emitted luminescence of the internal reference compound at wavelength λ 1 , said emitted luminescence at a wavelength λ 2 being modulated by the heavy atom effect; (e) calculating a corrected luminescence for the luminescence emitted by the fluorescent tracer compound at λ 2 based on the luminescence emitted by the internal reference compound at λ 1 ; and (f) correlating the corrected luminescence reading to the presence or amount of the target analyte in the sample.
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