Strategies to Enrich, Identify, and Characterize Glycoproteome in Blood Plasma Using Liquid Chromatography High-Resolution Mass Spectrometry.

Abstract

Blood and blood-derived components such as plasma and serum are considered as excellent resources that can be utilized to understand the biology of higher eukaryotic organisms including human beings. In research and clinical studies, blood plasma and serum are used to monitor health conditions, progression, and severity of diseases. Many of the disease-related studies utilize plasma and serum due to their disease relevance and accessibility as they can be collected from patients and healthy donors through minimally invasive approaches. Despite its significance, the unique proteome composition, complexity, wide dynamic range, and heterogeneity of the plasma proteins highlight critical factors that challenge the existing analytical technologies. Depletion of high abundant proteins is one among the accepted methods that can simplify the plasma proteome complexity; however, collateral loss of critical proteins should be anticipated. Selective protein enrichment seems to be a better alternative to depletion. Glycosylation of proteins is a dominant posttranslational modification known for its biological as well as diagnostic and therapeutic potential. Most of the reported therapeutic targets for diagnosis and monitoring are found to be glycosylated. In this chapter, a protocol for selective and reproducible enrichment of glycoproteins from blood plasma followed by identification through liquid chromatography high-resolution mass spectrometry has been documented.