Molecular Light Technology Limited

University College Cardiff Consultants Limited

Richard Charles Brown

Ian Weeks

Peter Kille

Andrew Paul Morby

A method is provided for determining in a medium the presence or amount of an analyte which is capable of binding to a ligand partner to form a ligand complex, which method comprises: (a) contacting the medium with either: (i) a ligand partner conjugated with an enzyme being capable of catalysing a reaction, or one or more reactions in a cascade thereof, to produce hydrogen peroxide, or (ii) a ligand partner and either a competing analyte or an analog of said analyte, a competing analyte or analyte analog being capable of forming a ligand complex with the ligand partner and the competing analyte or analyte analog being conjugated with an enzyme capable of catalysing a reaction, or one or more reactions in a cascade thereof, to produce hydrogen peroxide, (b) optionally separating complexed and uncomplexed enzyme conjugates, (c) causing or allowing the reaction or cascade of reactions to occur to produce hydrogen peroxide by contacting the complexed or uncomplexed enzyme conjugate with a corresponding enzyme substrate; (d) contacting hydrogen peroxide with a substance capable of exhibiting chemiluminescence in the presence of hydrogen peroxide selected from the group consisting of acridinium compounds and analogs thereof having a conjugate base with a pKa less then 9 to generate a chemiluminescent reaction, and (e) detecting the occurrence of said chemiluminescent reaction to determine the presence or amount of the analyte. Kits for use in connection with the method are also disclosed.

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Cardiff, United Kingdom

Richard Charles Brown