Sensitive Chemiluminescent Assay of Alkaline Phosphatase with a Device for High Yield of Hydrogen Peroxide in a Enzyme Cycling Reaction:Application to ELISA for Urinary FDP

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概要

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• We developed a chemiluminescent method for determining alkaline phosphatase (ALP) based on the following reaction ; hydrolysis of NADP (ALP substrate), NAD/NADH cycling reaction with alchohol dehydrogenase and 1-methoxy-5-methylphenaziniummethylsulfate (M-PMS) in the presence of superoxide dismutase (SOD), and subsequent luminol reaction with peroxidase. SOD resulted in about a 4 times higher yield of H<SUB>2</SUB>O<SUB>2</SUB> and its accumulation in the cycling reaction by immediate conversion of O<SUB>2</SUB><SUP>-</SUP> to H<SUB>2</SUB>O<SUB>2</SUB>.<BR>The determination limit of ALP activity assayed in a ten-minute cycling reaction system was similar to that estimated with 3-(2'-spiroadamantane)-4-methoxy-4-(3'-phosphoryloxy) phenyl-1, 2-dioxetane (AMPPD) reagent. This technique was applied to ELISA for sensitive determination of urinary FDP.

• Japan Society of Clinical Chemistryの論文

著者

• 森 時則

  東邦大学医学部大森病院臨床検査医学

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• Sensitive Chemiluminescent Assay of Alkaline Phosphatase with a Device for High Yield of Hydrogen Peroxide in a Enzyme Cycling Reaction:Application to ELISA for Urinary FDP

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