Muséum national d'Histoire naturelle et Centre National de la Recherche Scientifique, UMR 7245, 57 rue Cuvier, 75231 Paris Cedex 05, France.
UCD Centre for Nanomedicine, School of Chemistry and Chemical Biology, University College of Dublin, Dublin, Ireland.
J Immunol Methods. 2014 Apr;406:124-30. doi: 10.1016/j.jim.2014.02.014. Epub 2014 Mar 6.
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.
微囊藻毒素（MCs）是由蓝藻产生的七肽毒素。它们在水生生态系统中的全球性存在促使开发了几种检测方法，包括基于抗体的方法。在这里，我们建议应用重组抗体技术来生产由所谓的 scFv 与大肠杆菌的碱性磷酸酶（AP）融合而成的双价比色免疫探针（scFv-AP）。重组抗体技术允许开发具有改进性能且适合检测 MCs 的特定探针。融合蛋白在重组细菌的周质中产生，并用于开发直接竞争酶免疫分析，用于特异性检测 MCs，而无需进一步纯化。该重组分子识别的表位被限定为所有 MCs 共有的一个基序。与化学交联抗体到比色酶相比，这种遗传方法具有许多优势，并且可能适用于水样分析和原位检测。
