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Related Concept Videos

Oligosaccharide Assembly01:24

Oligosaccharide Assembly

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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...
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Related Experiment Video

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Improved In-gel Reductive β-Elimination for Comprehensive O-linked and Sulfo-glycomics by Mass Spectrometry
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An improved method for galactosyl oligosaccharide characterization.

Sachin Patil1, Jeffrey Rohrer1

  • 1Thermo Fisher Scientific, 1214 Oakmead Parkway, Sunnyvale, CA 94085, USA.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|October 14, 2021
PubMed
Summary
This summary is machine-generated.

This study improves galactosyl oligosaccharides (GOS) characterization using advanced chromatography and mass spectrometry. The enhanced method provides detailed GOS composition analysis, aiding digestive and immune health research.

Keywords:
GOSHPAE-PADHRAMOrbitrap Mass SpectrometryPrebioticsQ Exactive

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Area of Science:

  • Carbohydrate chemistry
  • Analytical chemistry
  • Biochemistry

Background:

  • Galactosyl oligosaccharides (GOS) offer significant digestive and immune health benefits.
  • Enzymatic synthesis of GOS results in complex mixtures of varying sizes and linkages.
  • Accurate GOS characterization is crucial for understanding their biological functions.

Purpose of the Study:

  • To develop an improved analytical method for in-depth characterization of galactosyl oligosaccharides (GOS).
  • To enhance the resolution and identification capabilities for complex GOS mixtures.
  • To establish a foundation for a comprehensive database correlating GOS structure with elution behavior.

Main Methods:

  • High-performance anion-exchange chromatography (HPAE) with a CarboPac PA300 column (4 µm particle size).
  • Pulsed amperometric detection (PAD) coupled with Orbitrap mass spectrometry (MS).
  • Improved sample preparation for sodium removal to minimize peak dispersion before MS analysis.

Main Results:

  • Significantly improved resolution of oligosaccharides, particularly in the disaccharide region.
  • Routine identification of GOS with degrees of polymerization from 2 to 6 using mass spectra and MS/MS fragmentation.
  • Identification of 28 distinct oligosaccharides in a commercial GOS sample.
  • Initial correlation attempted between oligosaccharide structure and HPAE elution behavior.

Conclusions:

  • The combined HPAE-PAD-MS approach offers superior GOS characterization compared to existing methods.
  • This technique enables detailed analysis of GOS composition, crucial for health-related applications.
  • The study lays the groundwork for a valuable structure-elution behavior database for GOS analysis.