The glycan moieties attached to the monoclonal antibodies (mAbs) can directly affect their stability, bioactivity, and immunogenicity. Glycan analysis can profile the glycan structure and monitor the relative quantities of a particular set of glycans to ensure product quality. As a forward-looking company as well as a market leader in the field of antibody glycan analysis, Creative Biolabs has successfully developed a versatile technology platform to illustrate the structure and relative quantities of glycan in mAbs and screen the changes in glycosylation. Intact glycoprotein profiling is a good way to ascertain general glycan patterns and glycan heterogeneity.
Glycosylation is considered a critical quality attribute of therapeutic mAbs, which is vital in the determination of physicochemical properties (solubility, viscosity, protein folding, antigen binding, secretion, antigenicity, etc.) and biological functions [antibody-dependent-cell mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC)]. Analysis of released glycans and glycopeptides provides information about the composition and/or position of the glycan, while intact glycoprotein analysis offers complementary structural information and allows assignment of individual proteoforms. The analysis of intact glycoproteins is the most straightforward approach to providing a general overview on the glycome, the total glycosylation pattern of glycoproteins, glycolipids, or other types of biomolecules, and then to revealing the complexity of a glycoprotein sample.
With the development in mass spectrometry technology, intact glycoprotein analysis is more advanced, which is fast, robust, does not require enzymatic digestion (preventing undesired modifications induced by enzymatic treatments), and enables high-throughput analysis.
Fig.1 A representative workflow for mass spectrometry-based glycoproteomic analyses.1
Intact glycoproteins can be directly analyzed for general glycan patterns and glycan heterogeneity, which is widely used in the analysis of the entire repertoire of glycans attached to proteins. Moreover, it has the diagnostic potential for different stages of cancers and various other disease states, helping to discover new biomarkers. At Creative Biolabs, we can provide diverse technologies to perform intact glycoprotein analysis, including but not limited to:
Fig.2 Glycomics workflow by MALDI-TOF MS. (Jansen, 2016)
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References
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