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Colorimetric engineered immunoprobe for the detection and quantification of microcystins.
- Source :
-
Journal of immunological methods [J Immunol Methods] 2014 Apr; Vol. 406, pp. 124-30. Date of Electronic Publication: 2014 Mar 06. - Publication Year :
- 2014
-
Abstract
- Microcystins (MCs) are heptapeptide toxins produced by cyanobacteria. Their global occurrence in aquatic ecosystems has prompted the development of several detection methods, including antibody-based methods. Here, we propose to apply recombinant antibody technologies to the production of a bivalent colorimetric immunoprobe (scFv-AP) made of the so-called scFv fused to the alkaline phosphatase (AP) of Escherichia coli. Recombinant antibody technologies allow the development of specific probes with improved properties and suitable for the detection of MCs. The fusion protein was produced in the periplasm of recombinant bacteria and was used to develop a direct competitive enzyme immunoassay for specific detection of MCs without requiring further purification. The epitope recognized by the recombinant molecule was circumscribed to a motif common to all MCs. Such a genetic approach offers many advantages over chemical cross-linking of antibodies to colorimetric enzymes and may be adaptable to the analysis of water samples and in situ detection.<br /> (Copyright © 2014 Elsevier B.V. All rights reserved.)
- Subjects :
- Alkaline Phosphatase genetics
Coloring Agents
Epitopes immunology
Recombinant Proteins genetics
Single-Chain Antibodies genetics
Alkaline Phosphatase immunology
Colorimetry methods
Cyanobacteria isolation & purification
Enzyme-Linked Immunosorbent Assay methods
Escherichia coli Proteins immunology
Microcystins analysis
Recombinant Proteins immunology
Single-Chain Antibodies immunology
Water Microbiology
Subjects
Details
- Language :
- English
- ISSN :
- 1872-7905
- Volume :
- 406
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 24607607
- Full Text :
- https://doi.org/10.1016/j.jim.2014.02.014