Issue 24, 2011

Construction and interrogation of enzyme microarrays using scanning electrochemical microscopy – optimisation of adsorption and determination of enzymatic activity

Abstract

Scanning electrochemical microscopy (SECM) has been used to image and study the catalytic activity of horseradish peroxidase (HRP) immobilised in a patterned fashion onto glass slides. Microarrays of HRP islands could be deposited on amino-modified glass slides using glutaraldehyde crosslinking combined with the SECM being used as a micro-deposition device. The enzymatic activity of the immobilised enzyme on the surface was in the presence of its substrate observed to give rise to substantial positive feedback between the slide and the SECM microelectrode tip. Conversely when either blank slides – or slides coated with HRP which had been subsequently thermally denatured were utilised, these showed negative feedback effects. Various conditions such as enzyme concentration, incubation time and substrate concentration were systematically varied to optimise sensitivity. Regular arrays of HRP could be assembled and when imaged, displayed lower limits of detection of 1.2 × 10−12 mol ml−1 of benzoquinone.

Graphical abstract: Construction and interrogation of enzyme microarrays using scanning electrochemical microscopy – optimisation of adsorption and determination of enzymatic activity

Article information

Article type
Paper
Submitted
13 Jul 2011
Accepted
10 Oct 2011
First published
21 Oct 2011

Analyst, 2011,136, 5287-5293

Construction and interrogation of enzyme microarrays using scanning electrochemical microscopy – optimisation of adsorption and determination of enzymatic activity

W. S. Roberts, F. Davis, S. D. Collyer and S. P. J. Higson, Analyst, 2011, 136, 5287 DOI: 10.1039/C1AN15589J

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