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. 2019 Oct 29;9:15481. doi: 10.1038/s41598-019-50722-y

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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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D3-L11 recognizes HEL with high affinity. (a) Binding of VHH to immobilized HEL at 25 °C monitored by SPR. The sensorgrams corresponded to the signal obtained from two-fold dilution series between 2 nM (darkest blue) and 0.125 nM (lightest blue). The black traces corresponded to the best fit to a 1:1 binding model. (b) Energy diagram of the interaction of D3-L11 with HEL with respect to the unbound state. Left, center, and right, corresponded to the unbound, transition, and equilibrium states, respectively. Errors bars corresponded to the standard error. (c) Titration of D3-L11 with HEL by ITC. The top panel shows the injection data (red, baseline). The bottom panel corresponded to the binding isotherm, (red, non-linear fitting). The slope of the binding isotherm was too steep for a precise estimation of the dissociation constant. In contrast, this condition favors a precise determination of the change of enthalpy.