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. 2011 Nov 1;3(6):558–567. doi: 10.4161/mabs.3.6.18119

Figure 4.

Figure 4

ADCP using human IgG1 and IgG2 anti-CD142 intact mAbs and F(ab')2 fragments generated with IdeS. (A) Representative flow cytometry dot plots depicting GFP-expressing MDA-MB-231 cells and human macrophages detected with anti-CD11b and anti-CD14 mAbs conjugated to Alexa Fluor 647. The left plot depicts cells incubated in the absence of anti-CD142 mAbs and the plot on the right depicts cells incubated with 9 µg/ml human IgG1 intact anti-CD142 mAb. (B) Representative fluorescent microscopy images of macrophages and GFP-expressing MDA-MB-231 cells in the absence of mAb (left panel) and incubated with 9 µg/ml IgG1 intact anti-CD142 (right part). Macrophages were detected with anti-CD11b and anti-CD14 antibodies conjugated to Alexa Fluor 568 (red), whereas the green indicates GFP-expressing MDA-MB-231 cells. The white arrows indicate macrophages that had phagocytosed MDA-MB-231 cells. (C) Human macrophage-mediated ADCP activity against GFP-expressing cells using intact IgG1 anti-CD142 (black circles), IgG1 F(ab')2 of anti-CD142 (red squares), IgG2 anti-CD142 (blue up triangle), and IgG2 F(ab')2 of anti-CD142 (green down triangle) in the absence of human serum. (D) ADCP activity against GFP-expressing cells in the presence of 30% human serum from donor one (n = 2). (E) ADCP activity against GFP-expressing cells in the presence of 30% human serum from donor two (n = 2).