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. 2018 Dec 12;11(1):26–44. doi: 10.1080/19420862.2018.1550321

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© 2018 The Author(s). Published with license by Taylor & Francis Group, LLC

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.

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Figure 1. — Off-target binding analysis chip re-array assay.

After performing an array-based binding screen on 4,975 human receptors for SHR-1210-IgG1, confirmatory analyses of binding specificity were performed on chips in which plasmids encoding PD1 and putative SHR-1210 off-target binding proteins were arrayed and used to transfect HEK-293 cells. Effective transfection of all plasmids was confirmed by screening for the co-encoded marker ZS green (A). Separate chips were then probed in duplicate using SHR-1210-IgG1 (B), Isotype IgG1 (C), Rituximab (D), and no primary antibody (E). These analyses confirmed that only SHR-1210-IgG1 exhibited binding to PD1, but also exhibited unexpected off-target binding to VEGFR2, FZD5 and ULBP2 proteins.