High-throughput analysis of the plasma N-glycome by UHPLC

Adamczyk, Barbara,1985Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi,Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology (författare)

The understanding of glycosylation alterations in health and disease has evolved significantly and glycans are considered to be relevant biomarker candidates. High-throughput analytical technologies capable of generating high-quality, large-scale glycoprofiling data are in high demand. Here, we describe an automated sample preparation workflow and analysis of N-linked glycans from plasma samples using hydrophilic interaction liquid chromatography with fluorescence detection on an ultrahigh-performance liquid chromatography (UHPLC) instrument. Samples are prepared in 96-well plates and the workflow features rapid glycoprotein denaturation, enzymatic glycan release, glycan purification on solid-supported hydrazide, fluorescent labeling, and post-labeling cleanup with solid-phase extraction. The development of a novel approach for plasma N-glycan analysis and its implementation on a robotic platform significantly reduces the time required for sample preparation and minimizes technical variation. It is anticipated that the developed method will contribute to expanding high-throughput capabilities to analyze protein glycosylation. © Springer Science+Business Media New York 2017.

NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
Automation
Glycan analysis
High-throughput
N-linked glycosylation
Plasma
Robotics
Ultrahigh-performance liquid chroma-tography

Stöckmann, H. (författare)
O’Flaherty, R. (författare)
Karlsson, Niclas G.,1966Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi,Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology(Swepub:gu)xkanic (författare)
Rudd, P. M. (författare)
Göteborgs universitetInstitutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi (creator_code:org_t)

Ingår i:High-Throughput Glycomics and Glycoproteomics, Eds: Gordan Lauc & Manfred WuhrerNew York : Springer, s. 97-1081064-37459781493964918

Av författaren/redakt...: Adamczyk, Barbar ...; Stöckmann, H.; O’Flaherty, R.; Karlsson, Niclas ...; Rudd, P. M.

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