pr9b00303_si_001.pdf (826.66 kB)
A Rapid Array-Based Approach to N‑Glycan Profiling of Cultured Cells
journal contribution
posted on 2019-09-18, 21:15 authored by Peggi M. Angel, Janet Saunders, Cassandra L. Clift, Shai White-Gilbertson, Christina Voelkel-Johnson, Elizabeth Yeh, Anand Mehta, Richard R. DrakeTypically,
N-glycosylation studies done on cultured cells require
up to millions of cells followed by lengthy preparation to release,
isolate, and profile N-glycans. To overcome these
limitations, we report a rapid array-based workflow for profiling N-glycan signatures from cells, adapted from imaging mass
spectrometry used for on-tissue N-glycan profiling.
Using this approach, N-glycan profiles from a low-density
array of eight cell chambers could be reported within 4 h of completing
cell culture. Approaches are demonstrated that account for background N-glycans due to serum media. Normalization procedures are
shown. The method is robust and reproducible, requiring as few as
3000 cells per replicate with a 3–20% coefficient of variation
to capture label-free profiles of N-glycans. Quantification
by stable isotopic labeling of N-glycans in cell
culture is demonstrated and adds no additional time to preparation.
Utility of the method is demonstrated by measurement of N-glycan turnover rates due to induction of oxidative stress in human
primary aortic endothelial cells. The developed method and ancillary
tools serve as a foundational launching point for rapid profiling
of N-glycans ranging from high-density arrays down
to single cells in culture.