Scientific article
Open access

Quantitative N-Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis

Published inPharmaceutics, vol. 13, no. 11, 1744
Publication date2021-10-20
First online date2021-10-20

The identification and accurate quantitation of the various glycoforms contained in therapeutic monoclonal antibodies (mAbs) is one of the main analytical needs in the biopharmaceutical industry, and glycosylation represents a crucial critical quality attribute (CQA) that needs to be addressed. Currently, the reference method for performing such identification/quantitation consists of the release of the N-glycan moieties from the mAb, their labelling with a specific dye (e.g., 2-AB or RFMS) and their analysis by HILIC-FLD or HILIC-MS. In this contribution, the potential of a new cost- and time-effective analytical approach performed at the protein subunit level (middle-up) was investigated for quantitative purposes and compared with the reference methods. The robustness of the approach was first demonstrated by performing the relative quantification of the glycoforms related to a well characterized mAb, namely adalimumab. Then, the workflow was applied to various glyco-engineered mAb products (i.e., obinutuzumab, benralizumab and atezolizumab). Finally, the glycosylation pattern of infliximab (Remicade®) was assessed and compared to two of its commercially available biosimilars (Remsima® and Inflectra®). The middle-up analysis proved to provide accurate quantitation results and has the added potential to be used as multi-attribute monitoring method.

  • Glycan profiling
  • Quantitative analysis
  • Protein subunits
  • Monoclonal antibodies
Citation (ISO format)
DUIVELSHOF, Bastiaan et al. Quantitative N-Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis. In: Pharmaceutics, 2021, vol. 13, n° 11, p. 1744. doi: 10.3390/pharmaceutics13111744
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Article (Published version)
ISSN of the journal1999-4923

Technical informations

Creation11/15/2021 9:48:00 AM
First validation11/15/2021 9:48:00 AM
Update time03/16/2023 2:42:03 AM
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