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3. Glycosylation analysis

The current glycan analysis methods involve multiple and lengthy sample preparation steps which can affect the robustness of the analyses. The development of orthogonal, direct and simple method is therefore desirable. We suggest the use of FTIR spectroscopy as a suitable, rapid and powerful tool to analyse protein glycosylation. Two types of analyses can be realized using this tool:

  • Comparative study in terms of global glycosylation level (weight ratio between sugars and proteins): it has been demonstrated that the intensity of the spectral band related to carbohydrates (between 1200 and 900 cm-1) is correlated with the global glycosylation level.
  • Comparative study in terms of glycan composition: it has been shown that FTIR spectra of glycoproteins provide a global but accurate fingerprint of the glycosylation profile. This fingerprint is not only sensitive to large differences such as the presence or absence of several monosaccharides but also to smaller modifications of the glycan and monosaccharide content.

 

 

References:

Derenne, A.; Derfoufi, K.-M. ; Cowper, B.; Delporte, C.; Goormaghtigh, E. FTIR spectroscopy as an analytical tool to compare glycosylation in therapeutic monoclonal antibodies., Anal. Chim. Acta. 2020, 1112, 62–71.

Wiercigroch, E.; Szafraniec, E.; Czamara, K.; Pacia, M.Z.; Majzner, K.; Kochan, K.; Kaczor, A.; Baranska, M.; Malek, K. Raman and infrared spectroscopy of carbohydrates: A review. Spectrochim. Acta Part A Mol. Biomol. Spectrosc. 2017, 185, 317–335.

Khajehpour, M.; Dashnau, J.L.; Vanderkooi, J.M. Infrared spectroscopy used to evaluate glycosylation of proteins. Anal. Biochem. 2006, 348, 40–8.