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. 2022 Aug 8;219(9):e20220849. doi: 10.1084/jem.20220849

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© 2022 Hale et al.

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Figure S2. — BLI competition assays and SEC-MALS demonstrating molecular weight of IgM pentamer. (A) Intrapanel competition BLI plots displaying sensor-detected RBD binding after addition of the indicated IgG⁺ MBC-derived IgG mAb. (B) Competition with well-characterized mAbs and the IgM⁺ MBC-derived 204 IgG mAb for RBD by BLI. (C) SEC-MALS traces for the purified 297 IgM mAb. The light scattering trace is in blue, and the molecular weight (MW) calculated from the combination of scattering, refractive index, and UV absorbance is shown in red across the peak elution window. The average calculated MW is shown in the inset, along with the predicted MW (MW_pred). The error is based on the SD from the 56 scans along the peak elution profile; however, the expected uncertainty can be as large as 10% due to assumptions about molar extinction coefficients and glycan occupancy. The hydrodynamic radius (Rh) was estimated from dynamic light scattering, which was also measured online and indicated under the peak. (D) Competition of mAb 204 against the panel of eight IgG⁺ MBC-derived mAbs.