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Electrochemical flow-ELISA for rapid and sensitive determination of microcystin-LR using automated sequential injection system

Lebogang, Lesedi LU ; Jantra, Jongjit LU ; Hedström, Martin LU and Mattiasson, Bo LU (2017) In Sensors 17(7).
Abstract

An amperometric immunoanalysis system based on monoclonal antibodies immobilized on Sepharose beads and packed into a micro-immunocolumn was developed for the quantification of microcystin-LR. Microcystin-LR (MCLR) was used as a reference microcystin variant. Inside the immunocolumn, free microcystins and microcystin-horseradish peroxidase (tracer) were sequentially captured by the immobilized antibodies, and the detection was performed electrochemically using Super AquaBlue ELISA substrate 2,2 -azinobis(3-ethylbenzothiazoline-sulfonic acid) (ABTS). The ABTS•+ generated by enzymatic oxidation of ABTS was electrochemically determined at a carbon working electrode by applying a reduction potential set at 0.4 V... (More)

An amperometric immunoanalysis system based on monoclonal antibodies immobilized on Sepharose beads and packed into a micro-immunocolumn was developed for the quantification of microcystin-LR. Microcystin-LR (MCLR) was used as a reference microcystin variant. Inside the immunocolumn, free microcystins and microcystin-horseradish peroxidase (tracer) were sequentially captured by the immobilized antibodies, and the detection was performed electrochemically using Super AquaBlue ELISA substrate 2,2 -azinobis(3-ethylbenzothiazoline-sulfonic acid) (ABTS). The ABTS•+ generated by enzymatic oxidation of ABTS was electrochemically determined at a carbon working electrode by applying a reduction potential set at 0.4 V versus Ag/AgCl reference electrode. The peak current intensity was inversely proportional to the amount of analyte bound to the immunocolumn. The amperometric flow-ELISA system, which was automatically controlled through the CapSenze™ (Lund, Sweden) computer software, enabled determination of MCLR as low as 0.01 µg/L. The assay time was very short (20 min for one assay cycle). In addition, the electrochemical signals were not significantly affected by possible interferences which could be present in the real samples. Along with the simplicity of automation, this makes the developed method a promising tool for use in water quality assessment.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Amperometric biosensor, Detection, Flow-ELISA, Microcystin-LR, Sequential injection
in
Sensors
volume
17
issue
7
publisher
MDPI AG
external identifiers
  • scopus:85025150746
  • wos:000407517600178
ISSN
1424-8220
DOI
10.3390/s17071639
language
English
LU publication?
yes
id
3972ea2f-033d-41da-a341-0909c082ebf1
date added to LUP
2017-07-31 10:21:55
date last changed
2018-01-07 12:13:38
@article{3972ea2f-033d-41da-a341-0909c082ebf1,
  abstract     = {<p>An amperometric immunoanalysis system based on monoclonal antibodies immobilized on Sepharose beads and packed into a micro-immunocolumn was developed for the quantification of microcystin-LR. Microcystin-LR (MCLR) was used as a reference microcystin variant. Inside the immunocolumn, free microcystins and microcystin-horseradish peroxidase (tracer) were sequentially captured by the immobilized antibodies, and the detection was performed electrochemically using Super AquaBlue ELISA substrate 2,2<sup>′</sup> -azinobis(3-ethylbenzothiazoline-sulfonic acid) (ABTS). The ABTS<sup>•+</sup> generated by enzymatic oxidation of ABTS was electrochemically determined at a carbon working electrode by applying a reduction potential set at 0.4 V versus Ag/AgCl reference electrode. The peak current intensity was inversely proportional to the amount of analyte bound to the immunocolumn. The amperometric flow-ELISA system, which was automatically controlled through the CapSenze™ (Lund, Sweden) computer software, enabled determination of MCLR as low as 0.01 µg/L. The assay time was very short (20 min for one assay cycle). In addition, the electrochemical signals were not significantly affected by possible interferences which could be present in the real samples. Along with the simplicity of automation, this makes the developed method a promising tool for use in water quality assessment.</p>},
  articleno    = {1639},
  author       = {Lebogang, Lesedi and Jantra, Jongjit and Hedström, Martin and Mattiasson, Bo},
  issn         = {1424-8220},
  keyword      = {Amperometric biosensor,Detection,Flow-ELISA,Microcystin-LR,Sequential injection},
  language     = {eng},
  month        = {07},
  number       = {7},
  publisher    = {MDPI AG},
  series       = {Sensors},
  title        = {Electrochemical flow-ELISA for rapid and sensitive determination of microcystin-LR using automated sequential injection system},
  url          = {http://dx.doi.org/10.3390/s17071639},
  volume       = {17},
  year         = {2017},
}