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. Author manuscript; available in PMC: 2018 Aug 1.
Published in final edited form as: Nat Protoc. 2017 Jul 27;12(8):1702–1721. doi: 10.1038/nprot.2017.058

Figure 8.

Figure 8

SDS-PAGE and LC–MS characterization of glycoengineered Herceptin (5a–e) bearing non-natural N-glycans26. (a) SDS-PAGE analysis of 5a–e. Lane 0: marker, lane 1: commercial Herceptin, lane 2: Herceptin-Fucα1,6GlcNAc (2a), lanes 3–7, 5a–e; (b) LC–MS profiles of 5a. The multiple charged m/z data are labeled with charge numbers. The deconvolution mass spectrum is shown in the embedded box. (c) SDS-PAGE analysis of PNGase-F digestion of 5a. Lane 8: 5a, Lane 9: 5a after PNGase-F digestion; (d) MS profile of the released non-natural glycan (shown in glycan symbol) from 5a by PNGase-F digestion. The ion flow chromatography of the PNGase-F digested sample is shown in the embedded box, and the peak for the glycan is marked with an arrow.