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Structural identification of N-linked carbohydrates using the GUcal application: A tutorial.

Gabor Jarvas1, Marton Szigeti2, Andras Guttman1

  • 1Horváth Csaba Memorial Institute of Bioanalytical Research, University of Debrecen, Hungary; MTA-PE Translational Glycomics Research Group, University of Pannonia, Veszprém, Hungary.

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Analytical glycomics uses capillary electrophoresis with laser induced fluorescent detection (CE-LIF) to analyze biopharmaceuticals. The GUcal application simplifies glycan structure elucidation by automating glucose unit (GU) value calculations for CE-LIF data.

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Bioinformatics

Background:

  • Analytical glycomics plays a crucial role in characterizing biopharmaceuticals.
  • Capillary electrophoresis with laser induced fluorescent detection (CE-LIF) is a key technique for analyzing complex carbohydrates.
  • Existing bioinformatics tools for glycomics analysis are underdeveloped compared to instrumentation.

Purpose of the Study:

  • To introduce and demonstrate the practical application of the GUcal software for glycoinformatics.
  • To provide a tutorial on calculating glucose unit (GU) values from CE-LIF electropherograms.
  • To showcase the utility of GUcal in glycan structure elucidation of human immunoglobulin G.

Main Methods:

  • Utilized capillary electrophoresis with laser induced fluorescent detection (CE-LIF) for glycan separation.
  • Employed the GUcal application for automated calculation of glucose unit (GU) values.
  • Applied both the bracketing standard approach and the triple internal standard method within GUcal.
  • Performed glycan structure elucidation using a worked example involving human immunoglobulin G.

Main Results:

  • The GUcal application automates the calculation of GU values from CE-LIF electropherograms.
  • The software supports both the bracketing standard and triple internal standard methods for GU value determination.
  • A practical example demonstrated the efficient glycan structure elucidation of human immunoglobulin G using GUcal.
  • GUcal offers a simple, rapid, and effective solution for glycomics research and routine analysis.

Conclusions:

  • GUcal is a valuable glycoinformatics tool that streamlines the analysis of CE-LIF data.
  • The application facilitates accurate glycan structure identification through automated GU value calculation.
  • GUcal enhances the efficiency of both exploratory glycomics research and high-throughput routine analysis.