Fig. 2: Glutaminyl cyclase is involved in the binding of magrolimab to cancer cell lines. | Communications Biology

Fig. 2: Glutaminyl cyclase is involved in the binding of magrolimab to cancer cell lines.

From: Pyroglutamation of cell surface proteins CD47 and TRP1 by glutaminyl cyclase modulates therapeutic antibody binding

Fig. 2: Glutaminyl cyclase is involved in the binding of magrolimab to cancer cell lines.

a Titration of magrolimab (1:1 dilution, commencing at 100 ng/mL) followed by αhuman IgG clone QA19 PE (1:100) on A375, Daudi and Hap1 wildtype (blue) and QPCTL KO (red) cells. Data represents individual data point and the line is the mean percentage of MFI of magrolimab staining relative to the MFI of 100 ng/mL magrolimab on wildtype cells (n ≥ 3). b Titration of αCD47 clone 2D3 (1:1 dilution, commencing at 20 μg/mL) on A375, Daudi and Hap1 wildtype (blue) and QPCTL KO (red) cells. Data represents individual data point and the line is the mean percentage of MFI of αCD47 clone 2D3 staining relative to the MFI of 20 μg/mL αCD47 clone 2D3 on wildtype cells (n = 2). c Representative histograms of magrolimab (1:40,000) followed by αhuman clone QA19 PE (1:100), unstained (gray) or αhuman clone QA19 PE only (dark gray) staining on A375, Daudi and Hap1 cells treated with 10 μM glutaminyl cyclase inhibitors PQ912 (red) or SEN177 (orange) or equal amounts of DMSO (blue) for three days. Bar graphs show the mean MFI of magrolimab, + standard deviation error bar (n = 3). One-way Anova was performed followed by the Dunnett’s multiple comparisons test. d Representative histograms of magrolimab (1:40,000) followed by αhuman clone QA19 PE (1:100), unstained (gray) or αhuman clone QA19 only (dark gray) staining on Daudi wildtype (blue), CD47 KO (red) and CD47 KO cells transduced with the CD47 mutant (purple; n = 3). Abbreviations: MFI, median fluorescent intensity; KO, knockout; US, unstained; Sec, secondary antibody only; PQ, PQ912; SEN, SEN177.

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