Automatic and rapid identification of glycopeptides by nano-UPLC-LTQ-FT-MS and proteomic search engine.

摘要: Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and Byonic™ proteomic search engine for separation, detection, identification N- O-glycopeptide glycoforms in standard glycoproteins. The use a BEH C18 nanoACQUITY column allowed separation glycopeptides present glycoprotein digest baseline-resolution same glycopeptide on basis number sialic acids. Moreover, evaluated several acquisition strategies order to improve detection characterization with maximum percentages. proposed strategy is simple set up technology platforms commonly used labs. method allows straightforward rapid obtention general glycosylated map given protein, including glycosites their corresponding structures. MS selected this work, based gas phase fractionation approach, led 136 unique peptides from four proteins, which represented 78% total identified. does not require an extra enrichment step, thus preventing bias that step could cause towards certain species. Data are available via ProteomeXchange identifier PXD003578.