An integrated strategy for high-sensitive and multi-level glycoproteome analysis from low micrograms of protein samples

ABSTRACT: In this study, we aimed to develop a simple and integrated spintip-based glycoproteomics technology, termed Glyco-SISPROT, for challenging the lowest starting material limitation and achieving comprehensive view of glycoproteome with shortest sample processing time. By carefully integrating and optimizing SCX, C18 and Concanavalin A packing material and their combination in spintip format, both predigested peptides and protein lysates could be processed by Glyco-SISPROT with high efficiency. More importantly, deglycopeptide, intact glycopeptide and glycans released by multiple glycosidases could be readily collected from the same Glyco-SISPROT workflow for LC-MS analysis. To the best of our knowledge, this approach should be one of the most comprehensive glycoproteomic approach by using limited protein starting material. One significant benefit of Glyco-SISPROT is that whole processing time was dramatically reduced from a few days to only 6 hours with good reproducibility when protein lysates were directly processed by Glyco-SISPROT.

INSTRUMENT(S): Orbitrap Fusion, Q Exactive

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Ruijun Tian  

PROVIDER: MSV000083570 | MassIVE | Thu Mar 14 04:17:00 GMT 2019

REPOSITORIES: MassIVE