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. 2018 Jan 29;8:1760. doi: 10.1038/s41598-018-19784-2

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© The Author(s) 2018

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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Conjugation efficiency and biodistribution analysis of CRISPR-modified antibody. (a) Coomassie blue stained gel of sortase-mediated conjugation of modified antibody to GGG-fluorophore. (b) Fluorescence scan of the antibody-fluorophore, stained with anti-Flag-HRP, showing the antibody conjugated fluorophore (red tab) compared to free fluorophore (black tab). (c) HPLC fluorescence trace antibody-fluorophore conjugates at different molar ratios. (d) Quantitative analysis of the percent of antibody modification. (e) Binding of ¹²⁵I-labeled CRISPR-modified antibody to REN-ICAM and REN-WT cells. (f) Biodistribution of ¹²⁵I-labeled CRISPR-modified anti-ICAM mAb in mice at 30 min. Tissue uptake is indicated as mean ± SEM (n = 3). Biodistribution analysis was performed comparing ¹¹¹In-labeled site-specific CRISPR/Cas9- to chemically-modified mAb. (g) Biodistribution of ¹¹¹In-labeled anti-ICAM mAb modified in mice at 30 min. Tissue uptake is indicated as mean ± SEM. (h) Localization ratio of selected organs. Significant differences were determined by t-test with Bonferroni correction to account for multiple comparisons.