A Rapid Array-Based
Approach to N‑Glycan Profiling of Cultured
Cells
Posted on 2019-09-18 - 21:15
Typically,
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.
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Angel, Peggi M.; Saunders, Janet; Clift, Cassandra L.; White-Gilbertson, Shai; Voelkel-Johnson, Christina; Yeh, Elizabeth; et al. (2019). A Rapid Array-Based
Approach to N‑Glycan Profiling of Cultured
Cells. ACS Publications. Collection. https://doi.org/10.1021/acs.jproteome.9b00303
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AUTHORS (8)
PA
Peggi M. Angel
JS
Janet Saunders
CC
Cassandra L. Clift
SW
Shai White-Gilbertson
CV
Christina Voelkel-Johnson
EY
Elizabeth Yeh
AM
Anand Mehta
RD
Richard R. Drake