Oligosaccharide Analysis
Oligosaccharide Analysis - Related Videos
01:24
Oligosaccharide Assembly
0 Views •
2023
Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans. Multiple sugar molecules that may or may...
09:56
Hierarchical and Programmable One-Pot Oligosaccharide Synthesis
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2019
This protocol demonstrates how to use the Auto-CHO software for hierarchical and programmable one-pot synthesis of oligosaccharides. It also describes the general procedure for RRV determination experiments and one-pot glycosylation of SSEA-4.
10:17
Pulse-chase Analysis of N-linked Sugar Chains from Glycoproteins in Mammalian Cells
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Cited by 5 •
2010
We describe a method for analysis of the alteration of N-linked glycans through the early life of glycoproteins after their biosynthesis in mammalian cells. This is achieved by pulse-chase analysis of metabolically labeled glycans, enzymatic release from glycoproteins and examination by HPLC.
13:09
Mass Spectrometric Analysis of Glycosphingolipid Antigens
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Cited by 9 •
2013
A specific and sensitive method to gain insight into the expression profile of glycosphingolipid antigens in immune organs and cells is described. The method takes advantage of the ion trap mass spectrometry allowing step-wise fragmentation of glycosphingolipid molecules for structural analysis in comparison to chemically synthesized standards.
11:27
Analysis of SCAP N-glycosylation and Trafficking in Human Cells
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Cited by 17 •
2016
We describe a modified method for membrane fraction isolation from human cells and sample preparation for the detection of SCAP N-glycosylation and total protein by using western blot. We further introduce a GFP-labeling method to monitor SCAP trafficking using confocal microscopy. This protocol can be used in regular biology laboratories.
FAQs
Branching and glycosidic linkages determine how a carbohydrate chain is organized, so molecules with similar compositions can still have different structures and biological roles. Mapping these features helps explain differences in molecular recognition and function. This structural resolution is especially important when comparing cell-surface glycans, protein-linked carbohydrates, or disease-associated changes.
Liquid chromatography separates oligosaccharides before measurement, while mass spectrometry provides mass information that supports composition and structural characterization. Using both approaches reduces the chance that chemically different molecules will be treated as a single signal. Their combination therefore supports more informative comparisons of glycan profiles and helps connect measured structures with biological processes.
Enzymatic digestion and chemical derivatization provide additional structural information beyond an intact mass measurement. Digestion can probe features of the carbohydrate chain, whereas derivatization changes how structures are distinguished during analysis. These strategies help researchers discriminate isomers and identify glycosidic linkage patterns, improving interpretation when oligosaccharides share similar compositions or masses.
Releasing or isolating the carbohydrate chains separates them from their glycoconjugate context so their composition and structure can be examined more directly. This preparation step precedes chromatographic separation and mass spectrometric characterization in the described workflow. The resulting measurements can then be related back to the original biological source, such as a cell-surface glycan or glycosylated protein.
A typical workflow begins by releasing or isolating oligosaccharides from glycoconjugates. The recovered material is separated by liquid chromatography and analyzed by mass spectrometry to obtain mass and structural information. Researchers may add enzymatic digestion or chemical derivatization when isomeric structures or linkage patterns require further discrimination. The sequence of steps supports systematic structural comparison.
The method is useful when researchers need to connect carbohydrate structure with biological recognition or function. Applications described for biology include examining cell-surface glycans, protein glycosylation, host–pathogen interactions, and changes associated with disease or development. Because these contexts can involve structurally distinct carbohydrate patterns, detailed analysis provides information that broader molecular measurements may not resolve.
Structural data from oligosaccharide analysis can reveal carbohydrate patterns associated with disease or development, supporting biomarker discovery. The same information can guide glycoengineering by clarifying which glycan features are present and potentially functionally relevant. It also contributes to the development of carbohydrate-based therapeutics and diagnostics by linking molecular structure with biological recognition and measurable outcomes.