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. 2020 Apr 9;39(2):37–44. doi: 10.1089/mab.2020.0001

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© Yukinari Kato et al. 2020; Published by Mary Ann Liebert, Inc.

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

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FIG. 1. — Preparation of ADC P38B-DM1 using CCAP method. (A) The reaction of antibody with CCAP peptide generated the monovalent and divalent antibodies owing to the two modified sites of CCAP peptide on Fc. To remove the unmodified and the divalent-modified antibodies, the reaction mixture was subjected to to affinity chromatography conjugated with Z33 affinity peptide. (B) The modification of HSA with DM-1 was performed. (C) Sodium dodecyl sulfate–polyacrylamide gel electrophoresis of conjugate products under reducing conditions. Lane 1: molecular weight marker; Lane 2: Z33-azide monovalent modified P38B; Lane 3: Z33-azide monovalent modified P38B + DBCO-HSA-DM-1; Lane 4: Z33-azide monovalent modified P38Bf (core fucose-deficient version of P38B); Lane 5: Z33-azide monovalent-modified P38Bf + DBCO-HSA-DM-1; Lane 6: DBCO-HSA-DM-1. H: heavy chain, L: light chain. DBCO, dibenzocyclooctyne; ADC, antibody–drug conjugate; CCAP, chemical conjugation by affinity peptide; HSA, human serum albumin.