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. 2022 Jul 7;122(20):15865–15913. doi: 10.1021/acs.chemrev.1c01031

Figure 9.

Figure 9

Multiplexed sample preparation workflow for N- and O-glycan profiling. Intra- and interday repeatability of the optimized method. (A) Proteins are immobilized on a polyvinylidene fluoride (PVDF) membrane by the addition of a (pure) glycoprotein, cell lysates, or derived from biological material (e.g., plasma). N-Glycans are released by the addition of PNGase F and eluted from the PVDF membrane. The O-glycans are released by adding a release agent and eluted from the PVDF membrane followed by a labeling procedure (2-AB). Eventually, the samples were analyzed using C18 nano-LC-MS/MS followed by data analysis. (B) Inter- and intraday repeatability of the O-glycan workflow. Average relative intensities for the O-glycans are displayed for those with a relative abundance above 1% per day. Error bars represent the standard deviations. Graphics in (A) were created using https://biorender.com/: H, hexose; N, N-acetylhexosamine; F, fucose; S, N-acetylneuraminic acid. Reproduced with permission from ref (24). Copyright 2022 de Haan et al.