Flow amperometric detection of indigo for enzyme-linked immunosorbent assays with use of screen-printed electrodes |
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Authors: | Setsuko YajimaYuri Sonoyama Ken SuzukiKeiichi Kimura |
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Affiliation: | a Faculty of Systems Engineering, Wakayama University, Sakae-dani, Wakayama, 640-8510 Wakayama, Japan b Tsukuba Research Laboratory, NOF Corporation, Toukoudai, Tsukuba, 300-2635 Ibaraki, Japan |
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Abstract: | A simple and automated methodology for a sensitive electrochemical detection of enzyme immunoassays that employ alkaline phosphatase (AP) as label has been developed. A flow injection system with programmable pump, valve and cell functions, amperometric detection of indigo and screen-printed electrodes (SPEs) are responsible for the advantages of this methodology. Amperometric detection at a low potential of indigo, the product of the enzymatic hydrolysis of the substrate 3-indoxyl phosphate (IP), is combined with a flow injection system. This incorporates in the flow cell a disposable screen-printed board provided with a graphite working electrode. No electrode pretreatment is necessary to obtain reproducible signals. The system was applied to the determination by an enzyme-linked immunosorbent assays (ELISA) of pneumolysin (PLY), a toxin related to respiratory infections. Linear calibration curves for low and high concentration ranges were obtained. These were also performed in a proteic matrix and linearity was also obtained. |
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Keywords: | Flow electrochemical enzyme immunoassay Screen-printed electrodes ELISA Pneumolysin 3-Indoxyl phosphate Alkaline phosphatase |
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