A Rapid Array-Based Approach to N-Glycan Profiling of Cultured Cells

Research output: Contribution to journalArticle

Authors

  • Peggi M. Angel
  • Janet Saunders
  • Cassandra L. Clift
  • Shai White-Gilbertson
  • Christina Voelkel-Johnson
  • Elizabeth Yeh
  • Anand Mehta
  • Richard R. Drake

External Institution(s)

  • Medical University of South Carolina

Details

Original languageEnglish (US)
Pages (from-to)3630-3639
Number of pages10
JournalJournal of Proteome Research
Volume18
Issue number10
StatusPublished - Oct 4 2019
Peer-reviewedYes

Abstract

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.

    Research areas

  • N-glycan, N-glycosylation, array, imaging mass spectrometry, label-free, single cell, stable isotope

Citation formats

APA

Angel, P. M., Saunders, J., Clift, C. L., White-Gilbertson, S., Voelkel-Johnson, C., Yeh, E., ... Drake, R. R. (2019). A Rapid Array-Based Approach to N-Glycan Profiling of Cultured Cells. Journal of Proteome Research, 18(10), 3630-3639. https://doi.org/10.1021/acs.jproteome.9b00303

Harvard

Angel, PM, Saunders, J, Clift, CL, White-Gilbertson, S, Voelkel-Johnson, C, Yeh, E, Mehta, A & Drake, RR 2019, 'A Rapid Array-Based Approach to N-Glycan Profiling of Cultured Cells', Journal of Proteome Research, vol. 18, no. 10, pp. 3630-3639. https://doi.org/10.1021/acs.jproteome.9b00303