Direct MALDI Glycotyping of Glycoproteins toward Practical Subtyping of Biological Samples.

Abstract

The rapid analysis of glycan patterns (glycoforms) of glycoproteins can accelerate their quality control and biomarker discovery. We have focused on the direct analysis of glycoprotein glycoforms using matrix-assisted laser desorption/ionization in-source decay mass spectrometry (MALDI-ISDMS), called MALDI glycotyping. Our results show that the 1,5-diaminonaphthalene (DAN)/2,5-dihydroxybenzoic acid (DHB)/Na matrix can directly analyze the glycoforms in the femtomolar range of intact glycoproteins. The addition of DAN improved the morphology of the solid matrix due to the mixture of DAN and DHB, which significantly contribute to the high sensitivity of this direct analysis. Adding DAN significantly improved the sensitivity of the glycan precursor ions in the TOF/TOF analysis because of its enhanced fragmentation effect as an efficient UV-MALDI matrix. Further, practical glycoform analysis (glycotyping) of diluted biological samples containing glycoproteins, such as egg whites, was also successfully achieved.