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Fig. 1 | Journal of Hematology & Oncology

Fig. 1

From: A pH-dependent anti-CD47 antibody that selectively targets solid tumors and improves therapeutic efficacy and safety

Fig. 1

Generation and characterization of pH-dependent anti-CD47 antibodies. a Schematic diagram depicting the pH-dependent selection strategy. Library phages were binding to CD47-ECD in pH 6.0 buffer, and subsequently eluted by pH 7.4 buffer; enriched phages were rescued for the next round of panning or, single clones were selected for characterization. b Characterization of the pH-dependent binding of BC2 and BC27. Serial dilutions of antibodies (in phage-scFv form) binding to CD47-ECD at pH 7.4 and pH 6.0, measured by ELISA. c Representative SPR sensograms of pH-dependent antibodies binding to CD47-ECD at pH 7.4 and pH 6.8; CD47-ECD was diluted from the indicated starting concentrations. Both original sensorgrams (colored lines) and fitting curves (black lines) are shown. d Summary of SPR measurements of pH-dependent antibodies, and a non-pH-dependent control Hu5F9; the affinity measurement results (KD) shown represent the mean and standard deviation of three replicated experiments. pH-dependence is defined as the ratio of KD at pH 7.4 to pH 6.8. The position of the mutated residues and their corresponding sequences within the HCDR3 and LCDR3 of BC27 (and its variants) are shown. e Top, serially diluted antibodies binding to CD47-ECD from pH 6.2 to pH 7.6, measured by ELISA. Bottom, the calculated EC50 for the data of the top panel

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