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A rapid 2AB-UHPLC method based on magnetic beads extraction for N-glycan analysis of recombinant monoclonal antibody.
- Source :
-
Journal of Chromatography B: Analytical Technologies in the Biomedical & Life Sciences . Mar2022, Vol. 1192, pN.PAG-N.PAG. 1p. - Publication Year :
- 2022
-
Abstract
- • A UHPLC-FLD method for N-glycan analysis based on magnetic bead extraction and 2-AB labeling was established. • Three conventional pretreatment procedures for N-glycan analysis, including solid phase extraction and acetone precipitation methods, were compared using three different antibodies. • Our pretreatment method maximizes the retention of some low abundance oligosaccharides and shows a better precision. • This pretreatment method could provide a rapid and high-throughput workflow option for N-glycan analysis in biopharmaceutical industry. N-glycosylation is one of the major post-translational modifications, with significant effects on the mechanism of action, the efficacy, and the safety of antibody drugs or glycoproteins. With the growing application of therapeutic antibodies, routinely monitoring N-glycosylation becomes increasingly important during cell culture process development and quality control. However, the current pretreatment methods for N-glycan analysis are time- and labor-consuming. The purification procedure of enzymatically released glycans could also partly affect the accuracy of results due to its complexity. In this study, a rapid ultra-high performance liquid chromatography method based on magnetic bead extraction and 2-AB fluorescent labeling was developed and compared against three popular pretreatment methods for N-glycan profiling (two were solid phase extraction and the other was acetone precipitation). The method's repeatability results showed that magnetic bead extraction has higher precision (% relative standard deviation (RSD), 0.121.06%) than solid phase extraction (SPE) (%RSD, 0.38–8.02%) and acetone precipitation (%RSD, 0.42–8.58%). This robust pretreatment method also maximized the retention of some low abundance oligosaccharides, and may thus provide a rapid and high-throughput workflow option for N-Glycan analysis in the biopharmaceutical industry. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 15700232
- Volume :
- 1192
- Database :
- Academic Search Index
- Journal :
- Journal of Chromatography B: Analytical Technologies in the Biomedical & Life Sciences
- Publication Type :
- Academic Journal
- Accession number :
- 155259780
- Full Text :
- https://doi.org/10.1016/j.jchromb.2022.123139