High-mannose N-glycans affect the efficacy, pharmacokinetics, and stability of therapeutic antibodies. Therefore, detection of these glycans is an important part of quality control during therapeutic antibody production. However, the high hydrophilicity and inefficient ionization of high-mannose glycans make their detection difficult when using mass spectrometry. Derivatization of glycans to improve their ionization is time-consuming. This presentation will describe how controlled proteolysis using tryspin-containing membranes enhances detection of glycans on therapeutic antibodies. The digestion takes place within one minute and yields relatively large peptides to enable fast and sensitive detection of high-mannose glycans. Desalting using molecular-weight-cutoff filters also improves detection of these glycans compared to conventional C18-based desalting methods.

Enhanced detection of high-mannose glycans using controlled trypsin digestion and mass spectrometry

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