N-glycosylation analysis by HPAEC-PAD and mass spectrometry

Changes in protein glycosylation are a hallmark of most types of cancer including ovarian carcinoma. The structural elucidation of glycans is technically challenging and it requires complementary chromatographic and spectroscopic techniques among others. Here, we describe the profiling of N-glycans from glycoproteins of SKOV3 ovarian carcinoma cells by high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry (MALDI-TOF MS). Mass spectrometry as a complementary method enables precise mass determination of N-glycan mixtures thus corroborating data obtained from HPAEC-PAD mapping in conjunction with reference oligosaccharide structures.