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. 2019 Dec 20;12(1):9. doi: 10.3390/toxins12010009

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Dot-plots of the binding of single chain Fv (scFv) to botulinum neurotoxins (BoNT)/Hi. The ability of each yeast-displayed scFv to bind BoNT/Hi holotoxin was determined by flow cytometry. Binding of each scFv to BoNT/H LC-H_N is shown as a positive control. Binding was detected using mAb 6F5.4 [12] or 6H1 (for 74F1 and 74G6) and is shown on the y-axis. The level of scFv display is shown on the x-axis. The green gated population are those yeast that display scFv on their surface. Mean fluorescent intensity (MFI) values are shown within each dot-plot. The negative control MFI (yeast stained with secondary antibody only) was 94.2 ± 15.7. A test MFI > two standard deviations from control (>125.6) was considered positive for binding BoNT/H LC-HN or BoNT/Hi.