Comprehensive Glycan Analysis of Sphingolipids in Human Serum/Plasma

Methods Mol Biol. 2023:2613:289-299. doi: 10.1007/978-1-0716-2910-9_21.

Abstract

Glycosphingolipids (GSLs) are glycolipids with ceramide and carbohydrate head groups that play an important role in numerous biological processes. Previously, we performed GSL-glycan analysis of various cell lines and virus-infected cells using a glycoblotting approach. Recently, we developed several methods for sialic acid linkage-specific chemical modification to distinguish sialylated glycan isomers by mass spectrometry. In this chapter, we describe a method for analyzing GSL-glycans in human serum/plasma using glycoblotting combined with aminolysis-SALSA (sialic acid linkage-specific alkylamidation) and lactone-driven ester-to-amide derivatization (LEAD)-SALSA for comprehensive and detailed structural glycomics.

Keywords: Glycoblotting; Glycosphingolipid-glycan; Human plasma; Human serum; Lactone-driven ester-to-amide derivatization; Mass spectrometry; Sialic acid linkage-specific alkylamidation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Glycosphingolipids / metabolism
  • Humans
  • N-Acetylneuraminic Acid*
  • Polysaccharides / chemistry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
  • Sphingolipids*

Substances

  • Sphingolipids
  • N-Acetylneuraminic Acid
  • Glycosphingolipids
  • Polysaccharides