Open AccessCloning and plant‐based production of antibody MC 10E7 for a lateral flow immunoassay to detect [4‐arginine]microcystin in freshwater
Author(s) -
Melnik Stanislav,
Neumann AnnaCathrine,
Karongo Ryan,
Dirndorfer Sebastian,
Stübler Martin,
Ibl Verena,
Niessner Reinhard,
Knopp Dietmar,
Stoger Eva
Publication year - 2018
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12746
Subject(s) - immunoassay , dipstick , nicotiana benthamiana , biology , monoclonal antibody , antibody , microbiology and biotechnology , virology , biochemistry , immunology , urine , virus
Summary Antibody MC 10E7 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 MC 10E7 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.