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N33D mutation has significantly reduced antigen binding affinity and CDC effector function. (A) Biacore sensorgrams of 7.5 nM wild-type and N33D mutant binding to immobilized CD52 peptide mimic. (B and C) Binding sensorgrams and global fit using a 1:1 binding model to obtain the binding constants. For wild-type (panel B), 0.2–7.5 nM antibody were used; for N33D (panel C), 0.2–1920 nM antibody were used. (D) CDC assay to test the effector function of the N33D mutant.
