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Current approaches to glycoprotein analysis.

Andrew K Hagan, Maomao Wang, Li Liu¹

¹Glycomics and Glycan Bioengineering Research Center, College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang, Nanjing 210095, P.R. China. lichen.liu@njau.edu.cn.

Protein and Peptide Letters

|July 1, 2014

Summary

Characterizing complex glycoproteins, vital for therapeutics like cancer antibodies, remains challenging. This review details analytical techniques, including HPLC, CE, and MS, for glycoprotein analysis.

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Area of Science:

Biochemistry
Analytical Chemistry
Biotechnology

Background:

Glycoproteins are crucial in therapeutics, treating conditions from myocardial infarction to cancer.
Protein glycosylation (N- and O-linked) is a complex posttranslational modification vital for protein function.
Characterizing these complex structures is a significant analytical challenge.

Purpose of the Study:

To review current techniques for characterizing glycoproteins.
To focus on analytical methods applicable to therapeutic glycoproteins.
To highlight methods for oligosaccharide analysis.

Main Methods:

High-performance liquid chromatography (HPLC) for separation and analysis.
Capillary electrophoresis (CE) for high-resolution separation.
Mass spectrometry (MS) for structural elucidation and identification.
Chemical labeling methods for enhanced detection sensitivity.
Enzymatic and chemical methods for oligosaccharide release.
Exhaustive protein hydrolysis using non-specific proteases.

Main Results:

Detailed description of key analytical techniques (HPLC, CE, MS).
Discussion of chemical labeling strategies for sensitive glycoprotein detection.
Overview of established and modern methods for releasing oligosaccharides from proteins.
Focus on methods relevant to therapeutic glycoprotein characterization.

Conclusions:

Advanced analytical techniques are essential for characterizing complex therapeutic glycoproteins.
A combination of separation, detection, and release methods provides comprehensive glycoprotein analysis.
Accurate characterization is critical for the development and quality control of glycoprotein-based therapeutics.