Amperometric assay for aldolase activity: antibody-catalyzed ferrocenylamine formation

Analytical Chemistry
Amit SagiDoron Shabat

Abstract

Screening of new catalysts for aldolase activity is a major task in bioorganic chemistry. For this purpose, fast and convenient methods are required for the detection of the catalysts. We have developed the first amperometric assay for aldol or retro-aldol catalytic activity. A new ferrocene-aldol derivative was synthesized with redox activity significantly different from that of ferrocenylamine. It was shown that the reaction between aldolase antibody 38C2 and a ferrocene-aldol substrate generated free ferrocenylamine, which could be detected and quantified by simple electrochemical measurement. The amperometric assay was applied to perform a Michaelis-Menten analysis of catalytic antibody 38C2 in order to determine the enzymatic kinetic parameters.

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Citations

Oct 16, 2007·Analytical and Bioanalytical Chemistry·Bettina Seiwert, Uwe Karst
Mar 4, 2017·Organic & Biomolecular Chemistry·Sean GogginsChristopher G Frost
Nov 22, 2014·Chemical Communications : Chem Comm·Sean GogginsChristopher G Frost
Aug 23, 2017·Organic & Biomolecular Chemistry·Sam A SpringChristopher G Frost
Apr 28, 2016·The Analyst·Sean Goggins, Christopher G Frost
May 1, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sam A SpringChristopher G Frost

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