Instrument configuration for native N-linked oligosaccharide characterization by HPAE-PAD/MS
Provide application installation instructions for coupling HPAE-PAD analyses to the Thermo Scientific™ Q Exactive™ family of mass spectrometers. This technical note focuses on using this set up to profile N-linked glycans released from glycoproteins.
Preparation of peptide N-Glycosidase F digests for HPAE-PAD analysis
The work shown here describes glycoprotein deglycosylation with PNGase F, followed by an efficient sample pretreatment using desalting and detergent removal spin columns to remove high concentrations of salt and detergents from small-scale PNGase F digestions. The HPAE-PAD technique was used to separate and detect the oligosaccharides.
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This application note describes direct detection of native glycans as an alternative to the common techniques for glycan analysis that rely on derivatization reactions to render glycans detectable. The lack of a detectable chromophore in native glycans is overcome by using HPLC with charged aerosol detection, a detector that can quantitatively measure any non-volatile compound.
The goal of this study was to develop an HPAE-PAD method to profile the major neutral and charged asparagine-linked (N-linked) oligosaccharides from IgGs, including monoclonal antibodies.
In this study, the glycosylation of human transferrin (as a model of PSA) and PSA are investigated by HPAE-PAD using <10 µg of protein. Two methods are utilized to evaluate protein glycosylation. One method is used to investigate the N-linked oligosaccharides and glycan sialylation. A second method evaluates the potential
