RESUMEN
Glycosylation is a crucial posttranslational modification (PTM) that might affect the safety and efficacy of monoclonal antibodies (mAbs). Capillary electrophoresis-mass spectrometry (CE-MS) enables the characterization of the primary structure of mAbs. A bottom-up proteomic workflow is designed to provide detailed information about the glycosylation. In this chapter, we describe the validated experimental protocol applied for the characterization and relative quantification of mAbs N-glycosylation at the glycopeptide level.
Asunto(s)
Electroforesis Capilar , Glicoproteínas/análisis , Natalizumab/análisis , Procesamiento Proteico-Postraduccional , Espectrometría de Masa por Ionización de Electrospray , Espectrometría de Masas en Tándem , Glicosilación , Proyectos de Investigación , Flujo de TrabajoRESUMEN
Capillary electrophoresis-mass spectrometry (CE-MS) enables the characterization of the primary structure of ADCs. An analytical method based on a derived bottom-up proteomic workflow is designed to provide detailed information about the amino acid sequence, the glycosylation profiling, and the location on the peptide backbone of the conjugated drugs. Here we describe the experimental protocol applied on the characterization of cysteine-linked brentuximab vedotin (Adcetris®).