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Glycan play key roles in protein folding, cell-cell recognition, cancer metastasis, and the immune system. In cell, glycosylation comprise that many kinds of glycan structures associated with a specific glycoprotein have different functions. So, the biological meaning of glycosylation has made them a prime target for discovering biomarker in diseases. Identifying the numerous glycan, we have used new method to assess the diversity of the N-linked oligosaccharides without derivatization has been developed using on-line nano-liquid chromatography (nanoLC) and high resolution time-of-flight mass spectrometry. In three times MS/MS results, chips offered good sensitivity and reproducibility in separating a intact protein. In tandem mass spectra, glycan fragment ions were identified as N-glycan sequences for mining glycoproteome. As a result, we have identified several N-glycan structures from monoclonal antibody.