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Biotechnology and Applied Biochemistry (1999) 29, (67–72) (Printed in Great Britain)

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Amperometric biosensor involving covalent immobilization of choline oxidase and butyrylcholinesterase on a methacrylate–vinylene carbonate co-polymer

Lucio Doretti*¹, Daniela Ferrara†, Silvano Lora† and Giancarlo Palma‡

*Servizio di Prevenzione e Protezione del CNR, Corso Stati Uniti 4, 35127 Padova, Italy, †Istituto di Fotochimica e Radiazioni d'Alta Energia del CNR, Sezione di Legnaro, via Romea 4, 35020 Legnaro (Padova), Italy, and ‡Dipartimento di Chimica Fisica, Università di Padova, via Loredan 4, 35131 Padova, Italy

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Abbreviations used: BChE, butyrylcholinesterase; ChO, choline oxidase; HEMA, 2-hydroxyethylmethacrylate; VCA, vinylene carbonate.

¹ To whom correspondence should be addressed.

A two-enzyme sensor for the determination of choline esters was prepared by the covalent co-immobilization of choline oxidase and butyrylcholinesterase on polymer membranes, obtained by radiation-induced co-polymerization of 2-hydroxyethylmethacrylate and vinylene carbonate at low temperature. The enzyme-modified membrane was applied to a platinum electrode and the determination of substrates was based on the electrochemical detection of enzymically generated H₂O₂. The analytical characteristics of this sensor, including the optimization of immobilization procedures, calibration curves for different substrates, pH and temperature effects and stability, are described.

Received 7 July 1998/22 October 1998; accepted 27 October 1998

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