Filter-Aided N-Glycan Separation (FANGS): A Convenient Sample Preparation Method for Mass Spectrometric N-Glycan Profiling

Salina Abdul Rahman, Ed Bergström, Christopher J. Watson, Katherine M. Wilson, David A. Ashford, Jerry R. Thomas, Daniel Ungar*, Jane E. Thomas-Oates

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


We have developed a simple method for the release and isolation of glycoprotein N-glycans from whole-cell lysates using less than a million cells, for subsequent implementation with mass spectrometric analysis. Cellular protein extracts prepared using SDS solubilization were sequentially treated in a membrane filter device to ultimately release glycans enzymatically using PNGase F in the volatile buffer ammonium bicarbonate. The released glycans are recovered in the filtrate following centrifugation and typically permethylated prior to mass spectrometric analysis. We call our method "filter-aided N-glycan separation" and have successfully applied it to investigate N-glycan profiles of wild-type and mutant Chinese hamster ovary cells. This method is readily multiplexed and, because of the small numbers of cells needed, is compatible with the analysis of replicate samples to assess the true nature of glycan variability in tissue culture samples. © 2014 American Chemical Society.

Original languageEnglish
Pages (from-to)1167-1176
Number of pages10
JournalJournal of Proteome Research
Issue number3
Early online date22 Jan 2014
Publication statusPublished - 2014


  • cultured mammalian cells
  • filter-aided sample preparation
  • N -glycans
  • permethylation
  • SDS solubilization

Cite this