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Quantitative glycomics using liquid phase separations coupled to mass spectrometry.

Josh Smith1, Stefan Mittermayr2, Csaba Váradi2

  • 1National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Dublin, A94 X099, Ireland. jonathan.bones@nibrt.ie and School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, 152-160 Pearse Street, Dublin 2, D02 R590, Ireland.

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Analyzing protein glycosylation is crucial for understanding disease and developing biopharmaceuticals. This review details advanced liquid chromatography and mass spectrometry methods for accurately quantifying glycans, essential for drug quality and diagnostics.

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Area of Science:

  • Biochemistry and Molecular Biology
  • Analytical Chemistry
  • Biotechnology

Background:

  • Protein glycosylation, the attachment of glycans, significantly alters protein properties and is linked to cellular functions and disease states.
  • Glycoproteins are vital biopharmaceuticals, where glycosylation is a critical quality attribute affecting drug efficacy and safety.
  • The complex, non-templated assembly of glycans presents substantial analytical challenges for accurate characterization.

Purpose of the Study:

  • To critically review analytical quantitation approaches for released glycans.
  • To discuss methods applicable to biopharmaceutical and biomedical contexts.
  • To highlight challenges and solutions in glycan analysis.

Main Methods:

  • Liquid phase separations coupled with mass spectrometry for released glycans.
  • Exploration of sample derivatization strategies to enhance detectability and separation.
  • Discussion of various quantitation techniques including isotopic and isobaric labeling, and targeted analysis.

Main Results:

  • The review critically evaluates diverse analytical strategies for glycan quantitation.
  • It addresses key considerations such as sample preparation, ionization techniques, and labeling strategies for relative quantification.
  • The article emphasizes the necessity of advanced separation and detection methods due to glycan complexity and sample limitations.

Conclusions:

  • Accurate glycan quantitation is essential for biopharmaceutical quality control and disease diagnostics.
  • Advanced analytical techniques, particularly liquid chromatography-mass spectrometry, are indispensable for overcoming the challenges in glycan analysis.
  • Further development in analytical methodologies will enhance our understanding and application of glycosylation in medicine and biology.