Electrochemical flow-ELISA for rapid and sensitive determination of microcystin-LR using automated sequential injection system
(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
- Lebogang, Lesedi LU ; Jantra, Jongjit LU ; Hedström, Martin LU and Mattiasson, Bo LU
- organization
- publishing date
- 2017-07-16
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Amperometric biosensor, Detection, Flow-ELISA, Microcystin-LR, Sequential injection
- in
- Sensors
- volume
- 17
- issue
- 7
- article number
- 1639
- publisher
- MDPI AG
- external identifiers
-
- scopus:85025150746
- pmid:28714899
- 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
- 2024-01-28 22:50:35
@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>}},
author = {{Lebogang, Lesedi and Jantra, Jongjit and Hedström, Martin and Mattiasson, Bo}},
issn = {{1424-8220}},
keywords = {{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}},
doi = {{10.3390/s17071639}},
volume = {{17}},
year = {{2017}},
}