High-sensitivity profiling of glycoproteins from human blood serum through multiple-lectin affinity chromatography and liquid chromatography/tandem mass spectrometry

Factors intrinsic and extrinsic to blood hamper the development of a routine blood test for human prion diseases

“…Since glycoproteins constitute one of the major subproteomes of blood (Madera et al, 2007), and because aberrant alterations in protein glycosylation are associated with some neurodegenerative disease states such as CJD and Alzheimer's disease (AD) (Sáez-Valero et al, 2003;Silveyra et al, 2006), lectin-based diagnostic blood tests, which are able to detect glycosylation differences in the glycoproteins from TSE-infected and uninfected blood samples, have been evaluated by some research groups (Table 1). Lectins are a group of proteins that specifically bind to or cross-link carbohydrates on the surface of proteins, which makes it possible to quantify or identify different sugar types (Debray et al, 1981).…”

Tags for the Stable Isotopic Labeling of Carbohydrates and Quantitative Analysis by Mass Spectrometry

“…24,25 This work builds on knowledge obtained from proteomics into the design of tags utilizing chemistry appropriate for released glycans, compatible with tandem MS, and flexible to allow features such as chromophores and/or affinity handles. Deuterated tags were designed for use with widely accepted reductive amination chemistry 26, 27 and may later be modified for use with 13 C, 15 N isotopes or to incorporate affinity handles. The modules are connected with amide bonds that are less labile than glycosidic bonds and are unlikely to fragment under conditions used for glycan tandem MS.…”

Lectinomics I. Relevance of exogenous plant lectins in biomedical diagnostics

“…The applications, based on lectin methodologies encompasses not only the ability to detect the biomarker (the glycosylated antigen), but also to separate and characterize the composition of the glycosylation patterns (Kre unin et al 2007;Madera et al 2007;Plavina et al 2007;Wuhrer et al 2007;Xu et al 2007). Numerous diseases progress through several stages, which are accompanied by changes in glycosylation.…”

“…Many assays, that utilize lectins to identify glycoprotein markers in serum from cancer patients rely on a three steps procedure: lectin affinity selection, separation and characterization of the glycoprotein markers by mass spectrometry (Drake et al 2006;Madera et al 2007). Such methods were developed to determine extend of sialylated glycoproteins and N-linked glycosylation profiling in pancreatic cancer serum us-13 ing WGA, SNA, MAA (Zhao et al 2006) and enriched by Con A-affinity (Zhao et al 2007a).…”