Next Article in Journal
Effect of Environmental Factors on Intra-Specific Inhibitory Activity of Carnobacterium maltaromaticum
Next Article in Special Issue
Paralytic Shellfish Toxins and Cyanotoxins in the Mediterranean: New Data from Sardinia and Sicily (Italy)
Previous Article in Journal
The Skin Bacterium Propionibacterium acnes Employs Two Variants of Hyaluronate Lyase with Distinct Properties
Previous Article in Special Issue
Culturing Toxic Benthic Blooms: The Fate of Natural Biofilms in a Microcosm System
Article Menu
Issue 3 (September) cover image

Export Article

Open AccessArticle
Microorganisms 2017, 5(3), 58; doi:10.3390/microorganisms5030058

Rapid and Highly Sensitive Non-Competitive Immunoassay for Specific Detection of Nodularin

1
Molecular Biotechnology and Diagnostics, Department of Biochemistry, University of Turku, FI-20520 Turku, Finland
2
Marine Research Centre, Finnish Environment Institute, Hakuninmaantie 6, FI-00430 Helsinki, Finland
*
Author to whom correspondence should be addressed.
Received: 12 June 2017 / Revised: 27 August 2017 / Accepted: 5 September 2017 / Published: 12 September 2017
(This article belongs to the Special Issue Toxic Cyanobacteria and Toxic Dinoflagellates)
View Full-Text   |   Download PDF [1633 KB, uploaded 15 September 2017]   |  

Abstract

Nodularin (NOD) is a cyclic penta-peptide hepatotoxin mainly produced by Nodularia spumigena, reported from the brackish water bodies of various parts of the world. It can accumulate in the food chain and, for safety reasons, levels of NOD not only in water bodies but also in food matrices are of interest. Here, we report on a non-competitive immunoassay for the specific detection of NOD. A phage display technique was utilized to interrogate a synthetic antibody phage library for binders recognizing NOD bound to an anti-ADDA (3-Amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4(E),6(E)-dienoic acid) monoclonal antibody (Mab). One of the obtained immunocomplex binders, designated SA32C11, showed very high specificity towards nodularin-R (NOD-R) over to the tested 10 different microcystins (microcystin-LR, -dmLR, -RR, -dmRR, -YR, -LY, -LF, -LW, -LA, -WR). It was expressed in Escherichia coli as a single chain antibody fragment (scFv) fusion protein and used to establish a time-resolved fluorometry-based assay in combination with the anti-ADDA Mab. The detection limit (blank + 3SD) of the immunoassay, with a total assay time of 1 h 10 min, is 0.03 µg/L of NOD-R. This represents the most sensitive immunoassay method for the specific detection of NOD reported so far. The assay was tested for its performance to detect NOD using spiked (0.1 to 3 µg/L of NOD-R) water samples including brackish sea and coastal water and the recovery ranged from 79 to 127%. Furthermore, a panel of environmental samples, including water from different sources, fish and other marine tissue specimens, were analyzed for NOD using the assay. The assay has potential as a rapid screening tool for the analysis of a large number of water samples for the presence of NOD. It can also find applications in the analysis of the bioaccumulation of NOD in marine organisms and in the food chain. View Full-Text
Keywords: nodularin; cyanotoxin; hepatotoxin; non-competitive immunoassay; quantification of nodularin; anti-immunocomplex binder; synthetic antibody phage library nodularin; cyanotoxin; hepatotoxin; non-competitive immunoassay; quantification of nodularin; anti-immunocomplex binder; synthetic antibody phage library
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Supplementary material

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Akter, S.; Vehniäinen, M.; Kankaanpää, H.T.; Lamminmäki, U. Rapid and Highly Sensitive Non-Competitive Immunoassay for Specific Detection of Nodularin. Microorganisms 2017, 5, 58.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Microorganisms EISSN 2076-2607 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top