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Protocol for ultra performance liquid chromatography-mass spectrometry N-glycan analysis of the native and desialylated human apo- transferrin

This protocol describes the method used for N- glycan analysis of the native and desialylated human apo-transferrin. The protein N-glycans were released with the addition of 1.2 U of PNGase F and overnight incubation at 37°C. The released N-glycans were labeled with 2-AB and purified using hydrophilic interaction liquid chromatography solid-phase extraction (HILIC-SPE). Fluorescently labeled N-glycans were separated by UPLC using Waters BEH Glycan chromatography column. All glycan structures were annotated with MS/MS analysis using an ESI-QTOF-MS system

transferrin ; N-glycan analysis ; UPLC ; mass spectrometry

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