Rituximab variants RTX. SEFL1.1 or RTX.SEFL2.0 and negative control RTX.IgG2 have minimal CDC activity compared with the wild-type rituximab. Rituximab (RTX.IgG1), RTX.IgG2, RTX.SEFL1.1, and RTX.SEFL2.0 mAb at 1, 10, and 100 μg/ml were added to Raji lymphoma cells for 24 h and the percentage of cell killing measured by flow cytometric analysis after adding propidium iodide (5 μg/ml). Data points were collected in duplicate and percentage of propidium iodide-positive cells was measured against total gated cells.