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. 2017 Aug 22;7:9130. doi: 10.1038/s41598-017-07966-3

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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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Binding affinity of mbk6 and mbk11 as measured by SPR and ELISA. Panel a and b show the SPR sensorgrams of mbk6 and mbk11, respectively. SPR experiments were performed on a single cycle mode. CD16A was coated onto a CM5 chips and different concentrations of BiKEs was injected. The kinetics constants were obtained by fitting the sensorgrams. Panel c and d show binding to CD16A and gp140sc, respectively, measured by ELISA. 50 ng/well of CD16A-mFc and gp140sc were coated onto costar half area, high binding, polystyrene 96 wells plate and incubated with serially diluted BiKEs for 2 hours at room temperature, after extensive washing by PBS + 0.05% Tween-20, bound BiKEs were detected by using HRP conjugated anti-FALG antibody. Experiments were performed in triplicate and the error bars denote ± SD, n = 3.