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. 2019 Dec 19;12(1):1685350. doi: 10.1080/19420862.2019.1685350

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© 2019 Bristol Myers Squibb. Published with license by Taylor & 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 3. — Co-expression and purification of CD3/CD20 bispecific antibodies. Intact mass spectrometry of (a) CD3/CD20 mG1_KD D265A and (b) CD3/CD20 mG2a_H4 treated with PNGase overnight. CD3/CD20 mG2a_H4 was not completely deglycosylated, hence the presence of one or more G0F glycans in addition to the mass of the correct bispecific. (c) CD3/CD20 mG1_KD D265A and (d) CD3/CD20 mG2a_H4 were analyzed by SDS-PAGE in both non-reducing (lanes C-1 and D-1) and 50 mM dithiothreitol reducing (lanes C-3 and D-3) conditions. Asterisk (*) denotes glycosylated products, which run at a greater molecular weight. CD3/CD20 mG1_KD D265A and CD3/CD20 mG2a_H4 were also analyzed by SEC-MALS (e and f, respectively). High molecular weight (HMW) and low molecular weight (LMW) species were calculated as a percentage of the total area under the curve. Sedimentation coefficients for CD3/CD20 mG1_KD D265A and CD3/CD20 mG2a_H4 as measured by analytical ultracentrifugation (g and h, respectively).