Reversed-phase peptide mapping of glycoproteins using liquid chromatography/electrospray ionization-mass spectrometry.

Publisher Summary The complexity of carbohydrate structures mandates that a variety of analytical methods be used for the study of these forms. A promising new procedure is described in this chapter that uses high-performance liquid chromatography (HPLC), in conjunction with electrospray ionization mass spectrometry (ESI/MS), as a tool to identify the sites of glycosylation and the general nature of the glycosylation. ESI/MS can detect whether an oligosaccharide is O-linked or N-linked. It can also differentiate between complex, high-mannose, or hybrid forms. This chapter also explains the use of this approach to gain limited linkage order information, using collisioninduced dissociation (CID), with both a single- and triple-quadrupole mass spectrometer. While the approach of on-line MS mapping is a useful addition to the battery of techniques used to characterize carbohydrates it supplements, but does not replace other important methods, such as nuclear magnetic resonance spectroscopy (NMR), used to determine linkage positions or anomeric configurations.