Fast electrochemical detection of anti-HIV antibodies: Coupling allosteric enzymes and disk microelectrode arrays

Laczka, Olivier; Ferraz, Rosa María; Ferrer-Miralles, Neus; Villaverde, Antonio; Muñoz, Francesc Xavier; Campo, F. Javier del

doi:10.1016/j.aca.2009.03.008

Cited by 25 publications

(17 citation statements)

References 46 publications

(49 reference statements)

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“…As to the determination of β-Gal activity, since electrochemical determination was the most common method to detect the final product (PAP) in the enzymatic reaction [3], we used a commercial kit to conduct the electrochemical assay. The experimental procedure was completely the same with our method.…”

Section: Analysis By Conventional Methodsmentioning

confidence: 99%

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Measurement of enzyme activity using a plug-based electrochemical microdevice

et al. 2012

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Determination of the activity of an enzyme in a solution of nL order was conducted using an electrochemical device with a plug-based microfluidic processing function. β-galactosidase (β-Gal) and adenosine triphosphatase (ATPase) were used as analyte enzymes. β-Gal is often used to be conjugated with antibodies, whereas the latter plays critical roles in cell functions. The activity of the enzymes was determined by measuring the concentration of one of the final products, p-aminophenol (PAP) or hydrogen peroxide (H 2 O 2 ) by coulometry. Preparation of plugs of solutions of predetermined volumes and mixing of solutions was carried out using a Tjunction on the device. A partially shallow flow channel structure was designed in the sensing area to improve collection efficiency. The observed output charge was correlated well with the amount of PAP or H 2 O 2 , which served as an indicator of the enzyme activity.

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Section: Analysis By Conventional Methodsmentioning

confidence: 99%

“…Therefore, the substrate concentration should be at least 10 times the K m value. The K m value for this enzyme and substrate has been reported to be 0.43 mM [3]. Hence the final concentration of PAPG was set at 4.3 mM.…”

Section: B Determination Of the Activity Of β-Galmentioning

confidence: 99%

Measurement of enzyme activity using a plug-based electrochemical microdevice

et al. 2012

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“…In the experiments carried out in redox cycling mode, the collector electrode potential was set to 0 V vs. Ag/AgCl (3 M KCl). The substrate concentration (10 −3 mol L −1 ) and measure conditions had been previously optimized [39]. Data shown are the average from at least three independent measurements.…”

Section: Electrochemical Detection Ofˇ-galmentioning

confidence: 99%

Amperometric detection of Enterobacteriaceae in river water by measuring β-galactosidase activity at interdigitated microelectrode arrays

Laczka

García-Aljaro

Campo

et al. 2010

Analytica Chimica Acta

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“…The use of electrochemical immunosensor is supported by their selectivity, sensitivity, versatility and simplicity and short analysis time (Bange et al, 2005); they also are renowned for their potential for miniaturization, portability and low cost. They have been successfully used in the detection of proteins, antibodies, bacteria and whole cells (Ghindilis et al, 1998;Kim et al, 2006;Melanson, 2007;Khaled et al, 2008;Konstantinov et al, 2009;Laczka et al, 2009;Mata et al, 2010). Many different classical strategies are employed to try and mimic the architecture of classical immunoassays on electrode surfaces; the assembly of the immunosensor with the attachment of antibodies or antigens on the surface can be carried out by several methods such as polymer or sol-gel entrapment (Ramanavicius et al, 2010) covalent attachment (Hays et al, 2006) or physical adsorption (Díaz-González et al, 2005).…”

Section: Introductionmentioning

confidence: 99%