Glycosylation Analysis of Therapeutic Glycoproteins
Therapeutic monoclonal antibodies (mAbs) are glycoproteins produced by living cell systems. Glycosylation is one of the most common post-translational modifications (PTMs) of proteins. The glycan moieties attached to the proteins can directly affect protein stability, bioactivity, and immunogenicity. Therefore, glycan variants of a glycoprotein product must be adequately analyzed and controlled to ensure product quality. However, the inherent complexity of protein glycosylation, combining the vast diversity in glycan structure and composition add an additional level of daunting analytical challenge.
In the development of therapeutic glycoproteins (e.g., mAbs), especially biosimilar products, glycan analysis usually involves the use of complementary methods for assessing specific glycosylation attributes, including:
- glycosylation site
- glycan structure: the structure of the carbohydrate chains, the oligosaccharide pattern (antennary profile)
- abundance: the carbohydrate content (neutral sugars, amino sugars, and sialic acids)
Analytical methods:
| Methods | Application |
| High-performance liquid chromatography (HPLC) | Widely used to quantify the amounts of released oligosaccharides |
| Mass spectrometry (MS) | Coupled with HPLC remains a powerful tool in the characterization of glycosylation site(s) occupancy and carbohydrate structures |
| Lectin-based microarray | Directly measure glycan variations in an intact protein without the need of clipping glycans from the protein backbone |
| Capillary electrophoresis (CE) | Alone or in combination is commonly used to monitor heterogeneity in sialic acids on intact glycoproteins |
| Isoelectric focusing (IEF) | |
| Ion-exchange (IEX) |
The selection of appropriate methods for a specific glycoprotein will depend on the purpose of testing and the ability of individual method in analyzing intact glycoproteins, glycopeptides, released glycans, and monosaccharides.
We recently provide an update series of services and related products in glycan analysis for determining unique glycan features, including assessment of glycosylation site, glycan structure, and carbohydrate content.
Our experts apply our experience in designing programs to deliver the glycosylation information to meet the requirements of your development stage and strategic packages pertinent to your actual need. Our experience spans tailored services for glycan analysis and a variety of related products. We are ready to support your antibody product development and manufacture, helping you to meet all of your therapeutic glycoproteins characterisation needs.
Service Available:
A panel of services for the characterisation of glycoproteins that Creative Biolabs offers:
- Identification of glycosylation sites through LC-MS/MS peptide mapping
- Capillary electrophoresis methods for glycan profile
- Confirmation of glycan structural characterization (enzyme array, MALDI-TOF)
- Determination of glycosylation linkage (GC-MS)
- Isoform analysis via cIEF
- Determination of Gu or G0 values (CE or normal phase LC)
- Glycan occupancy measuring and compositional analysis
- Determination and quantitative analysis of sialic acid residues, total and specific
- LC-MS intact molecular weight for intact molecule heterogeneity analysis
- Quantitative analysis of total sugar (monosaccharides and oligosaccharide composition neutral and amino sugars)
- Glycosylation population profile analysis with glycan ID via LC-MS/MS
Products Available:
- Glycan Staining or Labeling
Sugar Oxidation and Aldehyde Reaction
Protein Gel Stains
- Glycoprotein Purification or Enrichment
| Cat No. | Product Name | Associated Applications | Price |
| Glyco-060CL | CBGlycoTM Glycoprotein Isolation Kit (Concanavilin A) | Isolation of glycoproteins | Inquiry |
| Glyco-061CL | Jacalin Agarose | Immobilized Jacalin (for human IgG purification) | Inquiry |
- Glycoproteome and Glycome Analysis by Mass Spectrometry
Proteases and Protein-Cleaving Reagents
SILAC Isotope Labeling Reagents and Kits
For research use only. Not intended for any clinical use.
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