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. 2019 Apr 2;9:5531. doi: 10.1038/s41598-019-41722-z

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Selected neoepitope specific monoclonal antibodies exclusively recognise the specific cleavage site of the corresponding BoNT. (a) Western blot results to compare detection of cleaved vs. uncleaved SNAP-25 or VAMP-2 by the Neo-mAbs. SNAP-25 or VAMP-2 was incubated with (+) or without (−) BoNT overnight at 37 °C in cleavage buffer, samples were loaded on two identical polyacrylamide gels of which one gel was used for Coomassie staining, and the second gel was applied for Western blotting using Neo-mAbs. (b) Endopep-ELISA demonstrating specific recognition of cleavage sites. VAMP-2 or SNAP-25 were coated on microtitre plates, cleaved with 10 ng/mL BoNT as indicated and cleavage products were detected with Neo-mAbs indicated above each panel. Results from two independent experiments, each performed in technical duplicates are shown (n = 4; mean ± SD).