High-throughput N-glycan screening method for therapeutic antibodies using a microchip-based DNA analyzer: a promising methodology for monitoring monoclonal antibody N-glycosylation

• Published in: Analytical and bioanalytical chemistry Vol. 413; no. 19; pp. 4727 - 4738
• Main Authors: Kinoshita, Mitsuhiro, Nakajima, Kazuki, Yamamoto, Sachio, Suzuki, Shigeo
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.08.2021; Springer; Springer Nature B.V
• Subjects: Analysis; Analytical Chemistry; Antibodies; Biochemistry; Biological properties; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Cost analysis; Demand analysis; Deoxyribonucleic acid; DNA; DNA sequencers; Electrophoresis; Food Science; Glycan; Glycoproteins; Glycosylation; Heterogeneity; High-throughput screening (Biochemical assaying); Identification and classification; Laboratory Medicine; Manufacturing; Manufacturing industry; Methodology; Methods; Monitoring/Environmental Analysis; Monoclonal antibodies; N-glycans; Paper in Forefront; Physicochemical properties; Polysaccharides; Properties; Quality management; Reagents; Sample preparation; Semiconductors; Workflow; Japan; glycosylation; High-throughput methodology; Microchip electrophoresis; Therapeutic antibodies
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-021-03434-0

N -Glycosylation of therapeutic antibodies is a critical quality attribute (CQA), and the micro-heterogeneity affects the biological and physicochemical properties of antibodies. Therefore, the profiling of N -glycans on antibodies is essential for controlling the manufacturing process and ensuring the efficacy and safety of the therapeutic antibodies. To monitor N -glycosylation in recombinant proteins, a high-throughput (HTP) methodology for glycan analysis is required to handle bulk samples in various stages of the manufacturing process. In this study, we focused on the HTP methodology for N -glycan analysis using a commercial microchip electrophoresis-based DNA analyzer and demonstrated the feasibility of the workflow consisting of sample preparation and electrophoretic separation. Even if there is a demand to analyze up to 96 samples, the present workflow can be completed in a day without expensive instruments and reagent kits for sample preparation, and it will be a promising methodology for cost-effective and facile HTP N -glycosylation analysis while optimizing the manufacturing process and development for therapeutic antibodies. Graphical abstract