Fast purification of glycans and glycopeptides using silk-packed micropipette tip for matrix-assisted laser desorption/ionization-mass spectrometry and high-performance liquid chromatography-fluorescence detection analysis

Kayili H. M., Barlas N., ATAKAY M., SALİH B.

MICROCHEMICAL JOURNAL, vol.139, pp.492-499, 2018 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 139
  • Publication Date: 2018
  • Doi Number: 10.1016/j.microc.2018.03.034
  • Journal Name: MICROCHEMICAL JOURNAL
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.492-499
  • Keywords: Glycopeptide enrichment, Glycan purification, HILIC, Mass spectrometry, Silk, HYDROPHILIC INTERACTION CHROMATOGRAPHY, PROTEIN GLYCOSYLATION ANALYSIS, EFFICIENT ENRICHMENT, GLYCOPROTEIN ENRICHMENT, MAGNETIC NANOPARTICLES, SELECTIVE ENRICHMENT, NATURAL BIOMATERIAL, CANCER BIOMARKERS, IMMUNE-SYSTEM, PROTEOMICS
  • Hacettepe University Affiliated: Yes

Abstract

A novel micropipette tip packed with a natural product generated from silk worms is introduced in this study for the specific separation and purification of glycopeptides and glycans. The enrichment and purification performances of the silk-packed micropipette tip were investigated for both glycopeptide and glycan levels. Site specific glycosylation analysis of tryptic IgG Fc glycopeptides by MALDI-MS was achieved using a protocol containing enrichment step with the silk-packed micropipette tip. The enrichment efficiency and specificity of the silk-packed pipette tip regarding to glycopeptides and glycans was evaluated using the digests of some glycoproteins having neutral and sialylated glycopeptides and glycans. The MS and HPLC-FLD results indicate that the silk packed micropipette tip has good adsorption capacity to glycoconjugates in HILIC enrichment mode. In addition, purification of 2-aminobenzoic acid labelled IgG, fetuin and human plasma N-glycans with silk-packed micropipette tip was performed, and thus salts (PBS), detergents (SDS, NP-40) and interfered solvents (such as DMSO etc.) were removed prior to MS and HPLC-FLD analyses. (C) 2018 Elsevier B.V. All rights reserved.