Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7609869 | Journal of Chromatography A | 2017 | 33 Pages |
Abstract
In recent years, two-dimensional liquid chromatography (2D-LC) has seen an enormous evolution and one of the fields where it is being widely adopted is in the analysis of therapeutic monoclonal antibodies (mAbs). We here further add to the many flavours of this powerful technology. Workflows based on heart-cutting (LC-LC) and comprehensive (LCÂ ÃÂ LC) 2D-LC are described that allow to guide the clone selection process in mAb and biosimilar development. Combining Protein A affinity chromatography in the first dimension with size exclusion (SEC), cation exchange (CEX) or reversed-phase liquid chromatography-mass spectrometry (RPLC-MS) in the second dimension simultaneously allows to assess mAb titer and critical structural aspects such as aggregation, fragmentation, charge heterogeneity, molecular weight (MW), amino acid sequence and glycosylation. Complementing the LC-LC measurements at intact protein level with LCÂ ÃÂ LC based peptide mapping provides the necessary information to make clear decisions on which clones to take further into development.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Koen Sandra, Mieke Steenbeke, Isabel Vandenheede, Gerd Vanhoenacker, Pat Sandra,