N-glycosylation is one of the most common and complex post-translational modification of proteins, which plays an important role in a variety of biological activities, i.e.receptor-ligand interaction, signal transduction and immune response.The global profiling of N-linked glycosylation sites is a prerequisite for the in-depth investigation on the biological functions of glycoproteins.However, traditional N-linked glycosylation site profiling protocol is usually performed off line, leading to some drawbacks such as long analysis time, resultant sample loss and manual manipulation.Therefore, the development of a fast and specific strategy to achieve high-throughput and high-sensitive glycoproteome analysis becomes indispensable.