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. 2017 Aug 29;7:9584. doi: 10.1038/s41598-017-09489-3

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© The Author(s) 2017

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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In vitro propagation of human GSS-associated misfolded rec-PrPs to human wild type recombinant proteins. GSS-associated misfolded rec-PrPs were subjected to serial rounds of recPMCA (R1–R10) in quadruplicate. A representative experiment for each round is shown after PK (25 µg/ml) digestion of the samples. Tubes containing wild type human rec-PrP (129M and 129V) without any seed were used as control for cross-contamination. Misfolded P102L-129M and A117V-129M rec-PrPs required just 3 to 5 rounds, respectively for an entire adaptation to the wild type protein. However, misfolded P102L-129V and A117V-129V rec-PrPs were unable to propagate over the wild type human 129V rec-PrP after 10 rounds of recPMCA. No PK-resistant signals in any of the unseeded samples suggests complete absence of cross-contamination. 3F4 monoclonal antibody (1:10,000) was used. Mw: Molecular weight.