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. 2018 Sep 4;8:13208. doi: 10.1038/s41598-018-31417-2

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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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Functional evaluation of the recombinant anti-CD20 mAb in vitro. (a,b) CD20-binding activity of the recombinant mAb. Daudi (a) and Raji (b) cells were incubated with serial log dilutions of the mAbs, and their binding activities were assayed by flow cytometry using a FITC-conjugated goat anti-human IgG (H + L). Data are presented as the mean ± SD (n = 3). (c,d) CDC induced by the anti-CD20 mAb. Daudi (c) and Raji (d) cells were incubated with increasing concentrations of mAbs in the presence of human serum. CDC activity was measured using a Luminescent Cell Viability Assay Kit. Data are presented as the mean ± SD (n = 3). (e) ADCC induced by the anti-CD20 mAb. ADCC activity against Raji cells was determined using human PBMCs as effector cells at an effector-to-target cell ratio of 5:1. The activity was measured using a standard LDH assay. Data are presented as the mean ± SD (n = 3).