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. 2014 Oct 30;6(5):1265–1273. doi: 10.4161/mabs.29825

Figure 2.

Figure 2.

2095–2 specificity on cell-bound F(ab’)2 fragments and ADCC/CDC restoration of function. (A) F(ab’)2 fragments of rituximab generated with IdeS (solid black circles), GluV8 (solid black up triangles), and MMP-3 (solid black squares) displayed comparable binding to WIL2-S cells. (B) The anti-hinge mAb 2095–2 bound to cells opsonized with IdeS-generated F(ab’)2 fragments (solid black circles), but not to GluV8-generated F(ab’)2 fragments (solid black up triangles) or MMP-3-generated (solid black squares). Data from (A) and (B) are representative of two independent experiments. (C) The anti-hinge mAb 2095–2 was capable of restoring ADCC function to IdeS-generated F(ab’)2 fragments (open up red triangles), but not to GluV8-generated F(ab’)2 fragments (open green diamonds) or MMP-3-generated F(ab’)2 fragments (open down blue triangles). Addition of 2095–2 alone (solid gray squares) or F(ab’)2 IdeS fragments (solid black up triangles), F(ab’)2 GluV8 fragments (solid black diamonds), or F(ab’)2 MMP-3 fragments (solid black down triangles) alone did not result in appreciable ADCC activity. Data are representative of 2 independent experiments (duplicate measurements per experiment). (D) The anti-hinge mAb 2095–2 was capable of restoring CDC function to anti-CD20 IdeS-generated F(ab’)2 fragments (open up red triangles), but not to GluV8-generated F(ab’)2 fragments (open green diamonds) or MMP-3-generated F(ab’)2 fragments (open down blue triangles). Addition of 2095–2 alone (solid gray squares) or F(ab’)2 IdeS fragments (solid black up triangles), F(ab’)2 GluV8 fragments (solid black diamonds), or F(ab’)2 MMP-3 fragments (solid black down triangles) alone did not result in appreciable CDC activity. Data are representative of 2 independent experiments (triplicate measurements per experiment).