Quantifying Carbohydrate–Protein Interactions by Electrospray Ionization Mass Spectrometry Analysis

The development of analytical methods capable of characterizing carbohydrate–protein interactions, which are critical for many biological processes, represents an active area of research. Recently, the direct electrospray ionization mass spectrometry (ESI-MS) assay has emerged as a valuable tool for identifying and quantifying carbohydrate–protein complexes in vitro. The assay boasts a number of strengths, including its simplicity, speed, low level of sample consumption, and the unique ability to directly probe binding stoichiometry and to measure multiple binding equilibria simultaneously. Here, we describe the implementation of the direct ESI-MS assay for the determination of carbohydrate–protein binding stoichiometries and affinities. Common sources of error encountered with direct ESI-MS analysis of carbohydrate–protein interactions are identified along with strategies for minimizing their effects. The application of ESI-MS and a catch-and-release strategy for carbohydrate library screening are also d...