Cloning and plant-based production of antibody MC10E7 for a lateral flow immunoassay to detect [4-arginine]microcystin in freshwater
- Author(s)
- Stanislav Melnik, Anna-Cathrine Neumann, Ryan Karongo, Sebastian Dirndorfer, Martin Stübler, Verena Ibl, Reinhard Niessner, Dietmar Knopp, Eva Stoger
- Abstract
Antibody MC10E7 is one of a small number of monoclonal antibodies that bind specifically to [Arg4]-microcystins, and it can be used to survey natural water sources and food samples for algal toxin contamination. However, the development of sensitive immunoassays in different test formats, particularly user-friendly tests for on-site analysis, requires a sensitive but also cost-effective antibody. The original version of MC10E7 was derived from a murine hybridoma, but we determined the sequence of the variable regions using the peptide mass-assisted cloning strategy and expressed a scFv (single-chain variable fragment) format of this antibody in yeast and a chimeric full-size version in leaves of Nicotiana tabacum and Nicotiana benthamiana to facilitate inexpensive and scalable production. The specific antigen-binding activity of the purified antibody was verified by surface plasmon resonance spectroscopy and ELISA, confirming the same binding specificity as its hybridoma-derived counterpart. The plant-derived antibody was used to design a lateral flow immunoassay (dipstick) for the sensitive detection of [Arg4]-microcystins at concentrations of 100-300 ng/L in freshwater samples collected at different sites. Plant-based production will likely reduce the cost of the antibody, currently the most expensive component of the dipstick immunoassay, and will allow the development of further antibody-based analytical devices and water purification adsorbents for the efficient removal of toxic contaminants.
- Organisation(s)
- External organisation(s)
- Universität für Bodenkultur Wien, Technische Universität München
- Journal
- Plant Biotechnology Journal
- Volume
- 16
- Pages
- 27-38
- ISSN
- 1467-7644
- DOI
- https://doi.org/10.1111/pbi.12746
- Publication date
- 06-2017
- Peer reviewed
- Yes
- Austrian Fields of Science 2012
- 106023 Molecular biology
- Keywords
- Portal url
- https://ucris.univie.ac.at/portal/en/publications/cloning-and-plantbased-production-of-antibody-mc10e7-for-a-lateral-flow-immunoassay-to-detect-4argininemicrocystin-in-freshwater(bed33327-9e45-4819-825e-021929a74a68).html