Comparing different domains of analysis for the characterisation of N-glycans on monoclonal antibodies

• Published in: Journal of pharmaceutical analysis Vol. 10; no. 1; pp. 23 - 34
• Main Authors: Carillo, Sara, Pérez-Robles, Raquel, Jakes, Craig, Ribeiro da Silva, Meire, Millán Martín, Silvia, Farrell, Amy, Navas, Natalia, Bones, Jonathan
• Format: Journal Article
• Language: English
• Published: China Elsevier B.V 01.02.2020; Xi'an Jiaotong University, Journal of Pharmaceutical Analysis; School of Chemical and Bioprocess Engineering, University College Dublin, Belfield, Dublin 4, D04 V1W8, Ireland; Characterisation and Comparability Laboratory, NIBRT-the National Institute for Bioprocessing Research and Training, Foster Avenue, Mount Merrion, Blackrock, Co. Dublin, A94 X099, Ireland%Department of Analytical Chemistry, Biohealth Research Institute (ibs.GRANADA), University of Granada, Granada, Spain%Characterisation and Comparability Laboratory, NIBRT-the National Institute for Bioprocessing Research and Training, Foster Avenue, Mount Merrion, Blackrock, Co. Dublin, A94 X099, Ireland; Xi'an Jiaotong University; Elsevier
• Subjects: Acids; Biopharmaceuticals; Chromatography; Constant region; Cytotoxicity; Drug development; Enzymes; Gas flow; Glycan analysis; Glycopeptide analysis; Glycosylation; Immunoglobulins; Intact mass analysis; Mass spectrometry; Methods; Monoclonal antibodies; N-Glycans; Native mass spectrometry; Peptide mapping; Peptides; Polysaccharides; Product safety; Proteins; Quality control; Scientific imaging; Structure-function relationships; Germany; Spain; Glycan analysis; N-Glycans; Glycopeptide analysis; Native mass spectrometry; Monoclonal antibodies; Intact mass analysis; Biopharmaceuticals; Mass spectrometry; Peptide mapping
• ISSN: 2095-1779; 2214-0883
• DOI: 10.1016/j.jpha.2019.11.008

With the size of the biopharmaceutical market exponentially increasing, there is an aligned growth in the importance of data-rich analyses, not only to assess drug product safety but also to assist drug development driven by the deeper understanding of structure/function relationships. In monoclonal antibodies, many functions are regulated by N-glycans present in the constant region of the heavy chains and their mechanisms of action are not completely known. The importance of their function focuses analytical research efforts on the development of robust, accurate and fast methods to support drug development and quality control. Released N-glycan analysis is considered as the gold standard for glycosylation characterisation; however, it is not the only method for quantitative analysis of glycoform heterogeneity. In this study, ten different analytical workflows for N-glycan analysis were compared using four monoclonal antibodies. While observing good comparability between the quantitative results generated, it was possible to appreciate the advantages and disadvantages of each technique and to summarise all the observations to guide the choice of the most appropriate analytical workflow according to application and the desired depth of data generated. [Display omitted] •Comparative analysis of N-glycans present on four monoclonal antibodies using ten analytical methods reported.•Data presented to allow for impartial selection of appropriate analytical strategy depending on depth of information desired and ease of approach.•Methods included intact and subunit strategies and glycopeptide characterisation compared to released N-glycan as the current gold standard.