Immunosensor for the determination of Staphylococcus aureus using a tyrosinase-mercaptopropionic acid modified electrode as an amperometric transducer

Analytical and Bioanalytical Chemistry
Vanessa Escamilla-GómezJosé Manuel Pingarrón

Abstract

An amperometric immunosensor for the quantification of Staphylococcus aureus based on the coimmobilization of rabbit immunoglobulin G (RbIgG) and tyrosinase on a mercaptopropionic acid self-assembled monolayer modified gold electrode is reported. A competitive mode in which protein-A-bearing S. aureus cells and antiRbIgG labeled with alkaline phosphatase (AP) compete for the binding sites of immobilized RbIgG was used. Monitoring of the affinity reaction was carried out by the amperometric detection at -0.15 V of phenol generated in the enzyme reaction with AP, at the tyrosinase-modified electrode through the electrochemical reduction of the o-quinone formed. Optimization of the working variables, such as the immunosensor composition and incubation times, the applied potential, the working pH and the concentration of phenyl phosphate used as the AP substrate, was carried out. Under the optimized conditions, both the repeatability of the measurements and the reproducibility of the responses obtained with different immunosensors yielded relative standard deviation values for the steady-state current lower than 10%. The immunosensor showed a dynamic range from 4.4x10(5) to 1.8x10(7) S. aureus cells mL(-1), with a detection limit o...Continue Reading

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Citations

Feb 1, 2012·Analytical and Bioanalytical Chemistry·Berta Esteban-Fernández de ÁvilaJosé M Pingarrón
Nov 26, 2008·Chemical Communications : Chem Comm·Shan HuangXiao Niu Peng
Jun 21, 2014·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·M H Fabres-KleinA O B Ribon
Mar 17, 2009·Biotechnology Journal·Fahriye Ceyda Dudak, Ismail Hakki Boyaci
Mar 31, 2019·Micromachines·Zhenguo ZhangXin Du

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