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EP0342024A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

Luminescent or luminometric assays

Assignee: Pfizer Health AB
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Filing: 1989-05-10
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Granted: —
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Expires: 2009-05-10
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Abstract

Abstract A luminescent or luminometric assay procedure involving a two-step reaction. In the first step a reaction takes place between a selected oxidant, an enzyme for catalyzing oxidation reactions of that oxidant, and a substrate for that enzyme, resulting in an intermediate product which is a sufficiently-powerful oxidizing agent to bring about luminescent oxidation of a chemiluminescent DPD. In the second step, that intermediate compound is reacted with DPD resulting in a brief and relatively intense emission of light. An assay kit to facilitate the carrying out of this assay procedure.

Inventors

  • 👤 Fred Ture Blomberg
  • 👤 Jan Ove Friberg
  • 👤 Jan-Olof Valdemar Glindre
  • 👤 Jari Juhani Kangasmetsä

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-11-03 18:45
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/542; G01N33/58; G01N33/581; G01N33/74

Claims Analysis

Total Claims: 15
Claim 1
Independent Claim 84 words
A luminescent or luminometric assay which comprises the following steps: (i) carrying out a reaction between a selected oxidant, an enzyme capable of catalyzing an oxidation reaction utilizing that oxidant, and an aromatic substrate for that enzyme, to obtain an intermediate product which comprises a sufficiently powerful oxidizing agent to cause luminescent oxidation of a chemiluminescent DPD; and (ii) carrying out a reaction between the intermediate product generated in the first stage and a chemiluminescent DPD, and detecting or measuring the luminescence produced thereby.
Claim 2
Dependent Claim 84 words
A luminescent or luminometric assay which comprises the following steps: (i) carrying out a reaction between a selected oxidant, an enzyme capable of catalyzing an oxidation reaction utilizing that oxidant, and an aromatic substrate for that enzyme, to obtain an intermediate product which comprises a sufficiently powerful oxidizing agent to cause luminescent oxidation of a chemiluminescent DPD; and (ii) carrying out a reaction between the intermediate product generated in the first stage and a chemiluminescent DPD, and detecting or measuring the luminescence produced thereby.
Claim 3
Dependent Claim 14 words
An assay according to Claim 1, in which said oxidising agent contains free radicals.
Claim 4
Dependent Claim 29 words
An assay according to Claim 1 or Claim 2, in which the substrate is an aromatic amine, a benzothiazole derivative, a phenyl derivative, a halophenol, or a substituted phenol.
Claim 5
Dependent Claim 20 words
An assay according to Claim 3, in which the substrate is TMBZ, NNNN-TMBZ, ABTS, luciferin, cinnamic acid, p-iodophenol, or p-phenylphenol.
Claim 6
Dependent Claim 21 words
An assay according to any preceding claim, in which the oxidant is a per compound, and the enzyme is a peroxidase.
Claim 7
Dependent Claim 14 words
An assay according to Claim 5, in which the per compound is hydrogen peroxide.
Claim 8
Dependent Claim 15 words
An assay according to Claim 5 or Claim 6, in which the peroxidase is HRP.
Claim 9
Independent Claim 28 words
An method according to any preceding claim, in which the enzyme is HRP and step (i) is carried out at a pH within the range 3.6 to 8.8.
Claim 10
Dependent Claim 22 words
An assay according to Claim 8, wherein the substrate is TMBZ and step (i) is carried out at a pH of 5.5.
Claim 11
Dependent Claim 23 words
An assay according to any preceding claim, in which step (ii) is carried out at a pH within the range 8.5 to 12.
Claim 12
Dependent Claim 26 words
An assay according to Claim 10, in which step (ii) is carried out at a pH within the range 10 to 11, and preferably about 10.6.
Claim 13
Dependent Claim 15 words
An assay according to any preceding claim, in which the DPD is luminol or isoluminol.
Claim 14
Dependent Claim 15 words
A kit for enabling an assay according to the present invention to be carried out.
Claim 15
Dependent Claim 15 words
A kit for enabling an assay according to the present invention to be carried out.

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