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. 2021 Apr 27;13(1):1914885. doi: 10.1080/19420862.2021.1914885

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© 2021 The Authors. Published with license byTaylor & Francis Group, LLC

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — LC/MS analysis of NTERM conjugate. (a) DAR profile of T-N-F. T-N-F was analyzed using Waters Synapt G-2 system after deglycosylation reaction. Species with DAR 0–DAR 7 were observed and marked at D0–D7. The apparent DAR is 3.2 that is calculated as the weighted-average of DAR. (b) Conjugation sites were identified by peptide mapping. Trypsin-treated trastuzumab (top) and T-N-F (down) fragments mixture were resolved using an UPLC system with ACQUITY UPLC PST (BEH) C18 column

LC, liquid chromatography; MS, mass spectroscopy; DAR, drug-to-antibody ratio; T-N-F, trastuzumab-MMAF conjugate synthesized via NTERM conjugation reaction; UPLC, ultra-performance liquid chromatography.