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. 2022 Dec 7;13:7554. doi: 10.1038/s41467-022-35276-4

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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. 5 — a Polyspecificity reagent (PSR) staining of yeast displaying D06 variants. For the moderately polyreactive D06 nanobody, 18 out of 21 variants that were computationally designed to decrease polyreactivity reduced levels of binding to insect cell PSR staining. Data in a comprise the mean +/− SEM of at least three independent experiments, each performed in technical triplicate. b PSR staining of yeast displaying E10’ variants. For the highly polyreactive E10’ nanobody, 15 out of 16 computationally predicted single and double substitutions reduced binding to PSR reagent. Data in b comprise the mean +/− SEM of at least three independent experiments, each performed in technical triplicate. Substitutions to CDRs 1, 2, and 3 are colored in blue, green, and orange.