METHODS FOR THE RAPID PREPARATION OF LABELED GLYCOSYLAMINES FROM COMPLEX MATRICES USING MOLECULAR WEIGHT CUT OFF FILTRATION AND ON-FILTER DEGLYCOSYLATION

摘要

Methods for preparing labeled glycosylamines from a complex matrix are provided. The methodology includes the steps of: denaturing glycoproteins in a complex matrix to form a denatured complex matrix mixture; loading the denatured complex matrix mixture onto a MWCO filtration device; adding a glycosidase enzymatic solution onto the MWCO filtration device to form a deglycosylated complex matrix mixture comprising glycosylamines; collecting glycosylamines released from the MWCO filtration device; and derivatizing glycosylamines with a rapid tagging reagent to form a plurality of labeled glycosylamines suitable for detection in various liquid chromatography systems and detectors.