Peroxidase Activity of DNA Aptamer–Pt Complexes Prepared with Cisplatin

Akon Higuchi Department of Chemical and Materials Engineering, National Central University, 300 Jhongda Road, Jhongli, Taoyuan 32001, Taiwan; Department of Reproduction, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan

Siou-Ting Yang Department of Chemical and Materials Engineering, National Central University, 300 Jhongda Road, Jhongli, Taoyuan 32001, Taiwan

Yi-Di Siao Department of Chemical and Materials Engineering, National Central University, 300 Jhongda Road, Jhongli, Taoyuan 32001, Taiwan

Pei-Vin Hsieh Department of Chemical and Materials Engineering, National Central University, 300 Jhongda Road, Jhongli, Taoyuan 32001, Taiwan

Hisashi Fukushima OneCell, Inc., Nagahama Bio-Incubation Center 6, 8-1281 Tamura, Nagahama, Shiga 526-0829, Japan

Yung Chang Department of Chemical Engineering, R&D Center for Membrane Technology, Chung Yuan Christian University, 200 Chung-Bei Road, Chungli, Taoyuan 320, Taiwan

Wen-Yih Chen Department of Chemical and Materials Engineering, National Central University, 300 Jhongda Road, Jhongli, Taoyuan 32001, Taiwan; Institute of Systems Biology and Bioinformatics, National Central University, 300 Jhongda Road, Jhongli, Taoyuan 32001, Taiwan

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Abstract

DNA aptamers carrying Pt nanoparticles prepared with cisplatin showed peroxidase enzymatic activity while retaining the specific binding ability of the aptamers. Optimal preparation conditions of DNA–Pt complex prepared with cisplatin were investigated on the synthesis at pH 7–11, a reaction time of 1–18 h and 90°C. The enzymatic reaction of DNA–Pt complex obeyed Michaelis–Menten kinetics. K _M for the DNA–Pt complex was found to be of the same order as K _M for hemin and hemin–DNA complex, but one order of magnitude higher than that of horseradish peroxidase. A sandwich type of DNA enzyme-linked aptamer assay (DLAA) using DNA–Pt complex successively detected target protein of thrombin. DLAA using DNA–Pt complex fractioned by ultrafiltration membranes having a molecular weight cut-off of 30 000 and 300 000 showed 1.9-times higher sensitivity than DLAA using DNA–Pt complex without fraction. The DNA–Pt complex having specific size was effective for the sensitive detection of thrombin in DLAA.

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Peroxidase Activity of DNA Aptamer–Pt Complexes Prepared with Cisplatin

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Peroxidase Activity of DNA Aptamer–Pt Complexes Prepared with Cisplatin

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