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Multi-site N-Glycan mapping study 2: UHPLC.

Ákos Szekrényes1, SungAe Suhr Park2, Eoin Cosgrave3

  • 1Horváth Csaba Laboratory of Bioseparation Sciences, University of Debrecen, Debrecen, Hungary.

Electrophoresis
|January 14, 2018
PubMed
Summary
This summary is machine-generated.

This study assessed ultra-high performance liquid chromatography (UHPLC) for N-glycan analysis precision across 12 international sites. Results confirm UHPLC platforms offer adequate reproducibility for general N-glycan analysis applications.

Keywords:
2-ABBiotherapeuticsIntercompany studyN-glycansUHPLC

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Glycomics

Background:

  • N-glycosylation analysis is crucial for biopharmaceutical development and quality control.
  • Standardized and reproducible analytical methods are essential for reliable N-glycan profiling.
  • Previous work evaluated capillary electrophoresis; this study focuses on UHPLC.

Purpose of the Study:

  • To evaluate the precision (repeatability and reproducibility) of N-glycosylation analysis using ultra-high performance liquid chromatography (UHPLC).
  • To assess the suitability of standard UHPLC platforms for general N-glycan analysis across multiple international sites.

Main Methods:

  • International collaborative study involving 12 participating sites.
  • Standardized assay using ultra-high performance liquid chromatography (UHPLC) with fluorescence detection.
  • Analysis of APTS-labeled N-glycans from a common sample lot, reagents, and columns.
  • Statistical analysis of elution time, peak area, and peak area percent data following ISO 5725-2 principles.

Main Results:

  • Demonstrated adequate reproducibility of N-glycan analysis within and across participating sites.
  • Elution time, peak area, and peak area percent values were consistently determined for peaks ≥0.1% peak area.
  • Statistical analysis confirmed the reliability of the UHPLC method.

Conclusions:

  • Standard UHPLC-based N-glycan analysis platforms are suitable for general use.
  • The method exhibits sufficient precision for routine application in N-glycan analysis.
  • This validates UHPLC as a robust technique for glycomic studies.