Cotton HILIC SPE Microtips for Microscale Purification and Enrichment of Glycans and Glycopeptides

Selman, M.H.J.; Hemayatkar, M.; Deelder, A.M.; Wuhrer, M. (2011)

Article / Letter to editor

Solid-phase extraction microtips are important devices in modern bioanalytics, as they allow miniaturized sample preparation for mass spectrometric analysis. Here we introduce the use of cotton wool for the preparation of filter-free HILIC SPE microtips. To this end, pieces of cotton wool pads (approximately 500 mu g) were packed into 10 mu L pipet tips. The performance of the tips was evaluated for microscale purification of tryptic IgG Fe N-glycopeptides. Cotton wool HILIC SPE microtips allowed the removal of salts, most nonglycosylated peptides, and detergents such as SDS from glycoconjugate samples. MALDI-TOF-MS glycopeptide profiles were very repeatable with different tips as well as reused tips, and very similar profiles were obtained with different brands of cotton wool pads. In addition, we used cotton HILIC microtips to purify N-glycans after N-glycosidase F treatment of IgG and transferrin followed by MALDI-TOF-MS detection. In conclusion, we establish cotton wool microtips for glycan and glycopeptide purification with subsequent mass spectrometric detection.

• hydrophilic-interaction chromatography sample preparation method tandem mass-spectrometry immunoglobulin-g human serum n-glycans glycosylation analysis identification glycoproteins proteins

• All authors: Selman, M.H.J.; Hemayatkar, M.; Deelder, A.M.; Wuhrer, M.
• Date: 2011-04-01
• Journal: Analytical Chemistry
• Volume: 83
• Issue: 7
• Pages: 2492 - 2499
• DOI: doi:10.1021/ac1027116