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. 2018 Sep 12;8:13654. doi: 10.1038/s41598-018-31856-x

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© The Author(s) 2018

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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Assessment of vaccine quality and immunogenicity following exposure to an elevated temperature. (a) Thermostability analysis of v2-SAT2 and v2-SAT2/O vaccines before (unheated) and after (heated) incubation at 45 °C for 2 hours. The respective temperatures of genome release, and hence capsid disassembly, for three independent experiments are shown in brackets. The reduction in fluorescence signal due to loss of intact capsid (146S) is represented by dashed and non-dashed arrows for heated v2-SAT2 and v2-SAT2/O, respectively. (b) Analysis of v2-SAT2 and v2-SAT2/O vaccines before and after incubation at 45 °C for 2 hours by double antibody sandwich ELISA using a single-domain llama antibody (M377F VHH) that specifically recognises 146S. Asterisks indicate a significant decrease (Tukey’s method; 95% confidence, P < 0.05) in the respective 146S:12S vaccine composition following heat treatment. (c,d) Groups of five calves were vaccinated with the respective vaccine at days 0 and boosted on day 21; blood samples were assayed at 0, 7, 14, 21, and 28 days post vaccination. Homologous and heterologous VNT results are shown. Error bars indicate standard deviation. (c) Mean virus neutralizing-antibody titers (VNT (log₁₀)) of calves vaccinated with heat-treated (45 °C for 2 hours) v2-SAT2 or v2-SAT2/O vaccines. Asterisks indicate a significant difference (Tukey’s method; 95% confidence, P < 0.05) in neutralising titres between vaccinate groups. (d) Mean virus neutralizing-antibody titers (VNT (log₁₀)) of calves vaccinated with unheated v2-SAT2 or v2-SAT2/O vaccines.