title: Cotton HILIC SPE microtips for microscale purification and enrichment of glycans and glycopeptides.
abstract: 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 μg) were packed into 10 μL pipet tips. The performance of the tips was evaluated for microscale purification of tryptic IgG Fc 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.
authors.affiliation: Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Maurice H J
authors.initials: MH
authors.lastname: Selman
authors.forename: Mahdi
authors.initials: M
authors.lastname: Hemayatkar
authors.forename: André M
authors.initials: AM
authors.lastname: Deelder
authors.forename: Manfred
authors.initials: M
authors.lastname: Wuhrer
mesh_terms: D000818:Animals; D002417:Cattle; D002845:Chromatography / Q000379:methods*; D032561:Cotton Fiber; D006020:Glycopeptides / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011134:Polysaccharides / Q000302:isolation & purification* / Q000378:metabolism; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*; D014357:Trypsin / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000378:metabolism
chemical_list: D006020:Glycopeptides; D007074:Immunoglobulin G; D011134:Polysaccharides; D000509:alpha-Fetoproteins; D014357:Trypsin; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase