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. 2021 Jun 7;11:12004. doi: 10.1038/s41598-021-91360-7

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Measurement of ADAs, representing anti-GX-G3 antibodies, in monkeys subjected to 4-week repeated subcutaneous injection with GX-G3 followed by a 4-week recovery period. Monkeys were separated into 4 injection subgroups: VC (vehicle control), T1 (1 mg/kg GX-G3), T2 (3 mg/kg GX-G3), and T3 (10 mg/kg GX-G3). ADA levels in monkey sera were measured using the method validated in the present study. (a) Immuno-competition assays were performed to discriminate between true and false positive samples. True positivity for GX-G3 specificity was identified based on a greater than 30% difference 30% in absorbance between the non-treated and GX-G3-treated samples. (b) Monkey serum samples determined to be true positives were prepared as serial three-fold dilutions beginning from 20-fold. The antibody titer of each sample was expressed as the highest fold-dilution displaying a positive response (absorbance value > normalized NCO value). (c) The mean antibody titer of each group was arithmetically calculated. VC, vehicle control group; D1, D8, D22 and D29, dosing days 1, 8, 22 and 29, respectively; R30, recovery day 30.