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. 2019 Aug 28;9:12466. doi: 10.1038/s41598-019-48907-6

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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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Agglutination assay for 3-PBA detection in the absence (A) and presence (B) of amilCP expression. Determination of 3-PBA hapten-BSA concentration necessary to induce agglutination of E. coli DH10B cells displaying anti-3-PBA VHH (VHH3-PBA, Top) and detection of 3-PBA in response to cell pellet formation at constant concentration of the conjugate (0.08 pmol with no amilCP co-expression and 0.04 pmol with amilCP coexpression) (Bottom). Red rectangles and circles represent regions in which cell agglutination occurred and the limit of detection, respectively. Pictures are a composite of photos that were taken every six wells per frame using a phone camera. This experiment was conducted in three biological replicates.