Rapid monitoring of high‐mannose glycans during cell culture process of therapeutic monoclonal antibodies using lectin affinity chromatography

Abstract: We developed a simple high‐performance liquid chromatography assay to monitor high‐mannose glycans in monoclonal antibodies by monitoring terminal alpha‐mannose as a surrogate marker. Analysis of glycan data of therapeutic monoclonal antibodies by 2‐aminobenzamide assay showed a linear relationship between high mannose and terminal mannose of Fc glycans. Concanavalin A has a strong affinity to alpha‐mannose in glycans of typical therapeutic monoclonal antibodies. To show that terminal mannose binds specifically to Concanavalin A column, exoglycosidase‐treated monoclonal antibodies were serially blended with untreated monoclonal antibodies. Linear responses of terminal‐mannose binding to the column and comparable data trending with high mannose levels by 2‐aminobenzamide assay confirmed that terminal‐mannose levels measured by the Concanavalin A column can be used as a surrogate for the prediction of high‐mannose levels in monoclonal antibodies. The assay offers a simple, fast, and specific capability for the prediction of high‐mannose content in samples compared with traditional glycan profiling by 2‐aminobenzamide or mass spectrometry‐based methods. When the Concanavalin A column was coupled with protein A column for purification of antibodies from cell culture samples in a fully automated two‐dimensional analysis, high‐mannose data could be relayed to the manufacturing team in less than 30 min, allowing near‐real‐time monitoring of high‐mannose levels in the cell culture process.

Object Details

• Location: Deutsche Nationalbibliothek Frankfurt am Main
• Extent: Online-Ressource
• Language: Englisch

References and Relationships

• Bibliographic citation: Rapid monitoring of high‐mannose glycans during cell culture process of therapeutic monoclonal antibodies using lectin affinity chromatography ; day:03 ; month:02 ; year:2022 ; extent:9 ; Journal of separation science ; (03.02.2022) (gesamt 9)

Contributors, Places and Time

• Creator: Kim, Jun ; Albarghouthi, Methal

Further information

• DOI: 10.1002/jssc.202100903
• URN: urn:nbn:de:101:1-2022020414002859593278
• Rights: Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Last update: 15.08.2025, 7:34 AM CEST