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Related Experiment Video

Updated: Mar 23, 2026

A Quantitative Glycomics and Proteomics Combined Purification Strategy
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A Quantitative Glycomics and Proteomics Combined Purification Strategy.

Elizabeth S Hecht1, James P McCord1, David C Muddiman2

  • 1Department of Chemistry, North Carolina State University.

Journal of Visualized Experiments : Jove
|March 30, 2016
PubMed
Summary

A new filter-aided N-glycan separation (FANGS) method enables high-throughput, quantitative analysis of N-glycans and proteins from plasma. This unbiased approach simplifies complex biological sample analysis for biomarker discovery and disease understanding.

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Area of Science:

  • Biochemistry
  • Proteomics
  • Glycomics

Background:

  • Integrating multi-biomolecule data is crucial for understanding biological processes, disease, and developing treatments.
  • N-linked glycosylation is a key post-translational modification regulating vital cell functions.
  • Current N-glycan analysis methods are often resource-intensive and lack high-throughput capabilities.

Purpose of the Study:

  • To adapt and validate a filter-aided N-glycan separation (FANGS) protocol for plasma samples.
  • To develop a high-throughput, quantitative method for simultaneous glycomic and proteomic analysis.
  • To enable direct glycosylation site analysis with minimized peptide deamidation.

Main Methods:

  • Adapted Filter-Aided N-glycan Separation (FANGS) for plasma samples.
  • Coupled FANGS with glycan hydrazide labeling for quantitative LC-MS analysis.
  • Performed tandem, shotgun proteomics and glycosylation site analysis.
  • Compared FANGS with solid-phase extraction (SPE) for N-glycan purification and analysis.

Main Results:

  • FANGS-purified N-glycans showed comparable abundance and class distribution to SPE.
  • Variability between FANGS and SPE protocols was not statistically significant.
  • In-line proteomic data depth matched standard trypsin digests.
  • Minimized peptide deamidation allowed for direct glycosylation site identification.

Conclusions:

  • Developed a novel, in-line, high-throughput, filter-based protocol for quantitative glycomics and proteomics.
  • The FANGS method offers an unbiased and efficient alternative to traditional glycan analysis.
  • This approach facilitates integrated biomolecular analysis for biological and clinical insights.