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. 2021 Jul 28;23(9):939–950. doi: 10.1016/j.neo.2021.07.006

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© 2021 The Authors. Published by Elsevier Inc.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig 1 — Structure, characterization, and internalization activity of the humanized GPC1-ADC

A, Flow cytometric analysis of GPC1 expression. The shaded histogram profile indicates the isotype control, and the open histogram indicates the humanized anti-GPC1 antibody-staining results. B, Structure of the humanized GPC1-ADC consisting of the humanized anti-GPC1 antibody (clone T2) conjugated to the MMAE payload. C, BxPC-3 cells are incubated with unconjugated humanized anti-GPC1 mAb (blue closed circles) or humanized GPC1-ADC (red closed triangles). The mean fluorescence intensity at various concentrations is shown. D, Time-course analysis of the internalization activity of the antibody or ADC in BxPC-3 cells. The left panel shows unconjugated humanized anti-GPC1 mAb (clone T2); and the right panel, humanized GPC1-ADC. E, GPC1-ADC internalizes and locates in the lysosomes of BxPC-3 cells. Cell surface and intracellular GPC1 visualized on fluorescence microscopy. Green indicates the humanized anti-GPC1 antibody (clone T2) or humanized GPC1-ADC; red, the lysosomal marker LAMP-1; and blue, 4′,6-diamidino-2-phenylindole-stained DNA. Scale bar: 10 mm. (Color version of figure is available online)