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. 2017 Aug 21;7:8953. doi: 10.1038/s41598-017-09518-1

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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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Bioactivity and affinity with DR5-Fc comparison of TRAIL-Trimer, TRAIL-Fc and native TRAIL. (a) Bioactivity detection of TRAIL-Trimer, TRAIL-Fc and native TRAIL. The IC₅₀ value of TRAIL-Trimer, TRAIL-Fc and native TRAIL were assessed using a TRAIL sensitive cell line COLO205 on a Real-Time Cell Analysis (RTCA) system. The IC₅₀ value was obtained according to the dose-response cell index (CI) curve. The IC₅₀ value of TRAIL-Trimer, TRAIL-Fc and native TRAIL is 23.2 ng/mL, 260.8 µg/mL, and 6.7 ng/mL, respectively. The bottom graph displays IC₅₀ values when molar ratio-adjusted based on theoretical molecular weights for each protein. (b) Kinetic parameters of TRAIL-Trimer, TRAIL-Fc and native TRAIL binding to the soluble DR5-Fc fusion protein was assessed by biolayer interferometry measurements. The Super Streptavidin biosensor tips of the ForteBio Octet RED 96 were coated with biotinylated DR5-Fc. The biosensor tips were dipped in increasing concentrations gradient of the same molarity (15.4 nM–123.5 nM) to measure binding of TRAIL-Trimer, TRAIL-Fc and native TRAIL to DR5-Fc and subsequently moved to wells containing buffer (PBS) to measure dissociation rates. Detailed results are shown in Table 1.