Chemistry: analytical and immunological testing – Optical result – With fluorescence or luminescence
Patent
1996-07-11
1998-03-03
Snay, Jeffrey
Chemistry: analytical and immunological testing
Optical result
With fluorescence or luminescence
422 52, 250 36K, G01N 2169
Patent
active
057233420
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
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.
BACKGROUND AND PRIOR ART
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 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 is, hence, an object of the present invention to improve said known methods, especially with respect to the sensitivity of an analyte detection in combination with electrochemiluminescent procedures.
Fine subject matter of the invention is a method for measuring electrochemiluminescent phenomena in a solution or a solid phase contiguous with a solution, wherein the electrochemiluminescence is measured at a temperature of the solution and/or the solid phase, which is above the freezing point of the solution, but less than 25.degree. C.
Another aspect of the invention is a method for measuring electrochemiluminescent phenomena by generating electrochemiluminescence by applying a voltage to a working electrode. Before the electrochemiluminescence is generated, a potential between +400 and -400 mV, compared to an Ag/AgCl reference electrode, is applied to the working electrode.
Yet another aspect of the invention is a method for measuring electrochemiluminescent phenomena by generating electrochemiluminescence by applying a voltage to a working electrode. Electrochemiluminescence is generated by applying a voltage between the redox potential of the electrochemiluminescent system and +800 mV.
Yet another aspect of the invention is a method for 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 Thesit, C14-E09, Genapol, C8-E09, Plantaren.RTM., and octylglucoside or mixtures thereof.
Yet another aspect of the invention is a method for detecting an analyte by means of said methods, suitable reagents, and an apparatus for carrying out the first mentioned method.
The subject matter of the invention is a teaching based on the above listed prior. The fundamentals of electrochemiluminescent procedures are described in greater detail in the cited prior art documents. Instruments for measuring electrochemiluminescence comprise a measuring unit with a vessel for a reagent solution, at least two electrodes (a working e
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Grant, J. Hackh's Chemical Dictionary, 4th ed., McGraw-Hill Book Co., New York, 1972, see definition "temperature, room", p. 664, 1972.
Giesen Ursula
Hoyle Nicholas
Klemt Volker
Muller Gunter
Neumann Ulrich
Boehringer Mannheim GmbH
Snay Jeffrey
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