Chemistry: analytical and immunological testing – Optical result – With fluorescence or luminescence
Patent
1996-03-18
1997-10-14
Gorgos, Kathryn L.
Chemistry: analytical and immunological testing
Optical result
With fluorescence or luminescence
250361C, 422 52, G01N 2176
Patent
active
056771924
DESCRIPTION:
BRIEF SUMMARY
This application is the U.S. National Stage (filed under 35 U.S.C. 371) of International application No. PCT/EP 94/01328, filed 27, Apr. 1994.
Subject matter of the invention are methods for measuring electrochemiluminescent phenomena, methods for detecting an analyte using said methods, reagent solutions which can be used in said method and an apparatus particularly suitable for carrying out said method.
Methods for measuring electrochemiluminescent phenomena have been known for some years. Such methods make use of the ability of special metal complexes to achieve, by means of oxidation, an excited state from which they decay to the ground state, emitting electromagnetic radiation. Methods of this kind and suitable metal complexes are described in WO 86/02734, for example.
This technology has continually become more sophisticated. In WO 90/05296, an amine, preferably tripropylamine, which is a strong reducing agent when oxidized, is added to the test composition. The electrochemical reaction occurs in an electrolyte where the electrochemiluminescence (ECL) moiety, i.e. the metal complex able to emit electromagnetic radiation and the amine can be oxidized. The description mentions phosphate buffer at a pH of 6-9, preferably 7-7.5, as a suitable electrolyte in aqueous solution.
To increase the electromagnetic radiation, WO 90/05302 proposes to add Triton X-100 or Triton N-401, a detergent, to this test composition. WO 90/05411 describes an improved apparatus for measuring ECL.
Further, it has become possible to use the technology for detecting analytes by coupling electrochemiluminescent labels to analytes, analyte analogs or analyte-specific substances. The electrochemiluminescence was used to determine the quantity of analyte present. The description mentions in particular immunoassays where conventionally used labels are replaced by electrochemiluminescent labels.
Further improvements and applications of this technology are described in WO 87/06706, WO 89/04392, WO 89/10552, WO 89/10551, WO 90/05301, and WO 90/11511. The disclosures of these publications are assumed to be known.
It was, hence, an object of the present invention to improve said known methods, especially with respect to the sensitivity of the analyte detection in combination with electrochemiluminescence assay procedures.
Subject matter of the invention is a method of measuring electrochemical phenomena in a solution or a solid phase contiguous with the solution, wherein the solution is a detergent selected from the group consisting of fat alcohol ethoxylate, Plantaren, and octylglucoside or a mixture thereof.
Moreover, subject matter of the invention is also a method of detecting an analyte using said method and suitable reagents for carrying out said method.
The subject matter of the invention is a teaching based on the above listed prior art. The fundamentals of electrochemiluminescent procedures are described in greater detail in these prior art documents. Instruments for measuring electrochemiluminescence comprise a measuring unit with the container for a reagent solution, at least two electrodes (a working electrode and a counter electrode) which are in contact with the reagent solution during the measurement, and a detector to measure the light generated in the electrochemiluminescent process. Usually, an initial voltage (prepolarization) is first applied to the solution. Subsequently, this voltage is increased via the redox potential of a substance. e.g. an amine, contained in the solution. The so oxidized substance excites a material, e.g. certain ruthenium complexes, which are capable of producing chemiluminescence, to emit light. The amount of light received by a detector within a given time interval is a measure for the presence of the quantity of an electrochemiluminescent material. Provided the electrochemiluminescent material is a label for an analyte, an analyte analog or an analyte-specific substance, e.g. in an immunoassay, the light received is a measure for the presence of the analyte.
Experience has shown that the comm
REFERENCES:
patent: 4927769 (1990-05-01), Chang et al.
H. Karatani et al, "Electrogenerated Chemiluminescence of Cyclic Hydrazides in an Alkaline Brij35 Micellar System" Journal of Photochemistry and Photobiology A: Chemistry, 54 No month available (1990) 311-319.
Giesen Ursula
Hoyle Nicholas
Klemt Volker
Muller Gunter
Neumann Ulrich
Boehringer Mannheim GmbH
Gorgos Kathryn L.
Starsiak Jr. John S.
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