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Quantitative detection of Microcystis aeruginosa (cyanobacteria) in water using single domain antibodies (VHH) : dissertationGbenga Folorunsho Oginni, 2024, doctoral dissertation
Abstract: Microcystis aeruginosa accumulation in freshwater poses a significant threat to aquatic organisms and human health. The toxicity of Cyanobacteria metabolites urges for the development of methods for their rapid and efficient detection but what is still almost completely missing is the availability of reagents for the quantification of M. aeruginosa cells in water to monitor the fluctuations of its population. In this study, nanobodies against cell surface antigens of the toxic Cyanobacteria M. aeruginosa were recovered bywhole-cell panning of a naive phage display library. Six unique sequences were identified and three of them sub-cloned and purified as fusion immunoreagents together with either green fluorescent protein or Avi-Tag to be used for diagnostics. Theirspecificity and sensitivity were evaluated by immunofluorescence, by fluorescent and colorimetric cell ELISA and by thermal lens spectrometry (TLS). No cross-reactivity with unrelated microalgae was detected, and both ELISA and TLS methods provided a linear range of detection of several logs. The limit-of-detection of TLS was as low as 1 cell/ml.
Keywords: cyanobacteria, nanobodies, phage display, thermal lens spectrometry, dissertations
Published in RUNG: 10.09.2024; Views: 796; Downloads: 12
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