Understanding the Interplay Between a Permanent Charge and Hydrophobicity on the Electrospray Ionization of Glycans and the Application of Hydrazide Hydrophobic Tagging Reagents Toward the Assay of N-linked Glycans

摘要

An optimization of the glycan derivatization reaction by hydrazone formation has been presented along with the analysis of reagent properties beneficial to ESI-MS detection. A permanent charge was shown to be detrimental to glycan analysis, and the application of a neutral hydrazide reagent to glycans cleaved 'in-gel' was demonstrated. In future studies, the ability to cleave glycans 'in-gel' will allow the glycans to be linked to the proteins from which they are cleaved. The majority of proteins will remain in the gel pieces and are proposed to be able to be used to determine the glycosylation of specific proteins by using proteomic bioinformatics methods. These studies linking glycosylation to specific proteins will prove invaluable to systems biology and give a broader perspective involving the roles and interactions of glycoproteins.