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. 2017 Aug 11;7:7896. doi: 10.1038/s41598-017-08333-y

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© The Author(s) 2017

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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Overall experimental design. (a) Schematic representation of a reference antibody intermediate (AI) sample (gray), and its modifications such as N-linked glycosylation (colors). (b) Schematic representation of a reference antibody drug conjugate (ADC). The ADC is an immunoglobulin G1-kappa monomethylauristatin E (MMAE) conjugate, where one or more of the inter-chain disulfide bridges of the AI were intentionally chemically modified to an average of three to four linker-drug-modified cysteine residues (red stars). The conjugation was not expected to impact post-translational modifications of the AI. More than 34 post-translational modifications were considered herein, and enriched minor variant samples were generated for some modifications. (c) Schematic representation of one of the forced degradation conditions (e.g., 3 mM AAPH) applied to AI. The forced degradation introduces modifications such as oxidation (orange rectangles). (d) Schematic representation of one of the forced degradation conditions (e.g., 3 mM AAPH) applied to ADC. As in AI, forced degradation introduces modifications such as oxidation (orange rectangles). The Fab (dark gray) and Fc (light gray) regions of the molecules are shown for clarity.