title: Colorimetric engineered immunoprobe for the detection and quantification of microcystins.
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.
authors.affiliation: Muséum national d'Histoire naturelle et Centre National de la Recherche Scientifique, UMR 7245, 57 rue Cuvier, 75231 Paris Cedex 05, France.
authors.forename: Larissa M
authors.initials: LM
authors.lastname: Alvarenga
authors.affiliation: UCD Centre for Nanomedicine, School of Chemistry and Chemical Biology, University College of Dublin, Dublin, Ireland.
authors.forename: Julien
authors.initials: J
authors.lastname: Muzard
authors.affiliation: Muséum national d'Histoire naturelle et Centre National de la Recherche Scientifique, UMR 7245, 57 rue Cuvier, 75231 Paris Cedex 05, France.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Ledreux
authors.affiliation: Muséum national d'Histoire naturelle et Centre National de la Recherche Scientifique, UMR 7245, 57 rue Cuvier, 75231 Paris Cedex 05, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Bernard
authors.affiliation: Muséum national d'Histoire naturelle et Centre National de la Recherche Scientifique, UMR 7245, 57 rue Cuvier, 75231 Paris Cedex 05, France; University Paris-Sud, School of Pharmacy, IPSIT, 5 rue J.-B. Clément, 92296 Châtenay-Malabry, France. Electronic address: billiald@mnhn.fr.
authors.forename: Philippe
authors.initials: P
authors.lastname: Billiald
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics / Q000276:immunology*; D003124:Colorimetry / Q000379:methods*; D004396:Coloring Agents; D000458:Cyanobacteria / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology; D029968:Escherichia coli Proteins / Q000276:immunology*; D052998:Microcystins / Q000032:analysis*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*; D014871:Water Microbiology
chemical_list: D004396:Coloring Agents; D000939:Epitopes; D029968:Escherichia coli Proteins; D052998:Microcystins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D000469:Alkaline Phosphatase
keywords: Cyanotoxin; Detection; Immunotracer; Microcystin; Quantification; scFv