title: Automated, high-throughput serum glycoprofiling platform.
abstract: Complex carbohydrates are rapidly becoming excellent biomarker candidates because of their high sensitivity to pathological changes. However, the discovery of clinical glycobiomarkers has been slow, due to the scarcity of high-throughput glycoanalytical workflows that allow rapid glycoprofiling of large clinical sample sets. To generate high-quality quantitative glycomics data in a high-throughput fashion, we have developed a robotized platform for rapid serum-based N-glycan sample preparation. The sample preparation workflow features a fully automated, rapid glycoprotein denaturation followed by sequential enzymatic glycan release, glycan purification on solid-supported hydrazide and fluorescent labelling. This allows accurate glycan quantitation by ultra-high performance liquid chromatography (UPLC). The sample preparation workflow was automated using an eight-channel Hamilton Robotics liquid handling workstation, allowing the preparation of almost 100 samples in 14 hours with excellent reproducibility and thus should greatly facilitate serum-based glyco-biomarker discovery.
authors.affiliation: NIBRT GlycoScience Group, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co.Dublin, Ireland. pauline.rudd@nibrt.ie.
authors.forename: H
authors.initials: H
authors.lastname: Stöckmann
authors.forename: R
authors.initials: R
authors.lastname: O'Flaherty
authors.forename: B
authors.initials: B
authors.lastname: Adamczyk
authors.forename: R
authors.initials: R
authors.lastname: Saldova
authors.forename: P M
authors.initials: PM
authors.lastname: Rudd
mesh_terms: D001774:Blood Chemical Analysis / Q000295:instrumentation*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D054794:Glycomics / Q000295:instrumentation*; D057166:High-Throughput Screening Assays / Q000295:instrumentation*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D011134:Polysaccharides / Q000097:blood*; D015203:Reproducibility of Results; D012371:Robotics / Q000295:instrumentation*; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000295:instrumentation
chemical_list: D011134:Polysaccharides