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

Sara Carillo; Raquel Pérez-Robles; Craig Jakes; Meire Ribeiro da Silva; Silvia Millán Martín; Amy Farrell; Natalia Navas; Jonathan Bones

doi:10.1016/j.jpha.2019.11.008

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

J Pharm Anal. 2020 Feb;10(1):23-34. doi: 10.1016/j.jpha.2019.11.008. Epub 2019 Nov 29.

Authors

Sara Carillo¹, Raquel Pérez-Robles², Craig Jakes^{1

3}, Meire Ribeiro da Silva¹, Silvia Millán Martín¹, Amy Farrell¹, Natalia Navas², Jonathan Bones^{1

3}

Affiliations

¹ Characterisation and Comparability Laboratory, NIBRT - the National Institute for Bioprocessing Research and Training, Foster Avenue, Mount Merrion, Blackrock, Co. Dublin, A94 X099, Ireland.
² Department of Analytical Chemistry, Biohealth Research Institute (ibs.GRANADA), University of Granada, Granada, Spain.
³ School of Chemical and Bioprocess Engineering, University College Dublin, Belfield, Dublin 4, D04 V1W8, Ireland.

Abstract

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.

Keywords: Biopharmaceuticals; Glycan analysis; Glycopeptide analysis; Intact mass analysis; Mass spectrometry; Monoclonal antibodies; N-Glycans; Native mass spectrometry; Peptide mapping.