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. 2022 May 23;2:56. doi: 10.1038/s43856-022-00113-8

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© The Author(s) 2022

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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Fig. 1 — Surface and ribbon views of the VHHs (the three CDRs are colored red, green, blue, respectively): A VHH-72 can bind the sensor in different orientations. B VHH-72-C13 is oriented on the sensor by the Cys link (yellow). The direct linkage to the gold electrode is shown rather than the thiol-PEG-MAL linker, considered more meaningful. C VHH11-D4-C13 is oriented on the sensor surface by the Cys link (yellow). D Ribbon views of the two VHHs bound to the Spike RBD (rainbow colored, white surface). Note the large distance between the two epitopes. VHHs affinity constants determined by BLI: VHH-72 (K_D = 36.6 nM), VHH-72-13C (K_D = 12.1 nM), VHH-H11D4-13C (K_D = 5 nM).