Is your protein correctly glycosylated?

Together with our clients, we’re expanding in the biopharmaceutical field to build a portfolio of methodologies for glycan analysis. Our aim is to provide the best combination of methods to suit your particular purpose.

When compared with O-linked glycosylation, the analysis of N-linked glycosylation is fairly straightforward, as scientists have been researching it for a longer period of time. The fastest, most accurate and most economical way is through the use of our mass spectrometers.

Questions you might have about glycosylation that can be answered by looking at N-linked glycans include “Is my protein glycosylated?”, “What do the glycans look like?” and “Are they helping or hindering the development of my drug?”

For example, you may like us to determine the structure and heterogeneity of glycosylation for an entire protein or for each potential glycosylation site. To determine the structure of N-glycans we’ll use mass spectrometry (MALDI or LC-ESI) after labelling or permethylation. But to determine the antennary of glycans, or to quantify glycans, we’ll use HPAEC-PAD. Being a typical HPLC method, it has the advantage that you then can easily validate the assay for release testing. The Normal-Phase HPLC (NP-HPLC) complements this setup by providing an additional, highly sensitive and high-resolution analysis of your samples.

You might also ask us to find the sites of glycan attachment. We will do this either by peptide mapping or MALDI-ISD.

Other information about a protein’s glycosylation can be combined with what we know about its N-linked glycans. For example, we can determine the composition and amount of glycans attached to a protein by monosaccharide analysis and sialic acid analysis. The structural heterogeneity of a glycoprotein can be visualized using 2D PAGE or intact mass.

The characterization of protein glycosylation is a requirement of ICH Q6B.

Technical information

N-linked glycans are a common feature of glycoproteins such as EPO, antibodies and FSH. They have been shown to be important in many ways. They have an influence on protein folding as well as other factors, such as protein solubility, stability, immune reactions and cell-to-cell interactions.

N-glycosylation increases the structural heterogeneity of proteins when a range of structurally similar - but not identical - glycans are attached to each glycosylation site.

Glycosylation may be directly involved in the formation of diseases, but also plays an important role in altering the pharmacokinetics by stabilizing protein conformation, improving solubility or protecting from proteases.

The N-glycosylation profile of a recombinant protein can vary largely depending on the organism chosen for expression.

Your contact for N-linked Glycosylation at Protagen Protein Services GmbH


Dr Marcus Mreyen

Director Business Development
Our business development team will be happy to assist you with your project.

Phone: +49 (0) 231 9742-6100