Electrochemical protease biosensor based on enhanced AC voltammetry using carbon nanofiber nanoelectrode arrays

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dc.contributor.author Swisher, Luxi Zhang
dc.contributor.author Syed, Lateef U.
dc.contributor.author Prior, Allan M.
dc.contributor.author Madiyar, Foram R.
dc.contributor.author Carlson, Kyle R.
dc.contributor.author Nguyen, Thu A.
dc.contributor.author Hua, Duy H.
dc.contributor.author Li, Jun
dc.date.accessioned 2013-07-29T19:50:45Z
dc.date.available 2013-07-29T19:50:45Z
dc.date.issued 2013-07-29
dc.identifier.uri http://hdl.handle.net/2097/16000
dc.description.abstract We report an electrochemical method for measuring the activity of proteases using nanoelectrode arrays (NEAs) fabricated with vertically aligned carbon nanofibers (VACNFs). The VACNFs of ~150 nm in diameter and 3 to 5 μm in length were grown on conductive substrates and encapsulated in SiO[subscript 2] matrix. After polishing and plasma etching, controlled VACNF tips are exposed to form an embedded VACNF NEA. Two types of tetrapeptides specific to cancer-mediated proteases legumain and cathepsin B are covalently attached to the exposed VACNF tip, with a ferrocene (Fc) moiety linked at the distal end. The redox signal of Fc can be measured with AC voltammetry (ACV) at ~1 kHz frequency on VACNF NEAs, showing distinct properties from macroscopic glassy carbon electrodes due to VACNF’s unique interior structure. The enhanced ACV properties enable the kinetic measurements of proteolytic cleavage of the surface-attached tetrapeptides by proteases, further validated with a fluorescence assay. The data can be analyzed with a heterogeneous Michaelis-Menten model, giving “specificity constant” k[subscript cat]/K[subscript m] as (4.3 ± 0.8) x 10[superscript 4] Mˉ¹sˉ¹ for cathepsin B and (1.13 ± 0.38) x 10[superscript 4] Mˉ¹sˉ¹ for legumain. This method could be developed as portable multiplex electronic techniques for rapid cancer diagnosis and treatment monitoring. en_US
dc.language.iso en_US en_US
dc.relation.uri http://pubs.acs.org/doi/abs/10.1021/jp312031u en_US
dc.rights Permission to archive granted by the editor of the Journal of Physical Chemistry C, March 28, 2013. This document is the unedited Author’s version of a Submitted Work that was subsequently accepted for publication in The Journal of Physical Chemistry C, copyright © American Chemical Society after peer review. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/jp312031u en_US
dc.subject Nanoelectrode array en_US
dc.subject Legumain en_US
dc.subject Cathepsin B en_US
dc.subject Enzymatic kinetics en_US
dc.subject AC voltammetry en_US
dc.title Electrochemical protease biosensor based on enhanced AC voltammetry using carbon nanofiber nanoelectrode arrays en_US
dc.type Article (author version) en_US
dc.date.published 2013 en_US
dc.citation.doi doi:10.1021/jp312031u en_US
dc.citation.epage 4277 en_US
dc.citation.issue 8 en_US
dc.citation.jtitle The Journal of Physical Chemistry C en_US
dc.citation.spage 4268 en_US
dc.citation.volume 117 en_US
dc.contributor.authoreid tanguyen en_US
dc.contributor.authoreid duy en_US
dc.contributor.authoreid junli en_US

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