mAbs were treated overnight with 4 U of PNGase F (Roche, IN, USA) at room temperature. Subsequently, treated and untreated samples were buffer-exchanged into 150 mM aqueous ammonium acetate (pH 7.5) by ultrafiltration (Vivaspin 500, Sartorius Stedim Biotech, Germany) with 30-kDa cut-off filter. Sample concentrations were adjusted to 5 μM before 4 μl was used for native mass spectrometry analysis. Native mass spectrometry analysis was carried out with a modified Exactive Plus Orbitrap instrument with extended mass range as described before (50). Intact Mass software was used for zero-charge deconvolution of the spectra to determine the antibody mass (51).

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