PMID- 27743361
OWN - NLM
STAT- MEDLINE
DCOM- 20180116
LR  - 20180412
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 1503
DP  - 2017
TI  - High-Throughput Analysis of the Plasma N-Glycome by UHPLC.
PG  - 97-108
AB  - 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.
FAU - Adamczyk, Barbara
AU  - Adamczyk B
AD  - Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, 
      Sahlgrenska Academy, University of Gothenburg, Box 440, 405 30, Gothenburg, 
      Sweden.
AD  - NIBRT GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research 
      and Training, Fosters Avenue, Mount Merrion, Blackrock, Co., Dublin, Ireland.
FAU - Stockmann, Henning
AU  - Stockmann H
AD  - NIBRT GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research 
      and Training, Fosters Avenue, Mount Merrion, Blackrock, Co., Dublin, Ireland.
AD  - AbbVie Inc., North Chicago, IL, USA.
FAU - O'Flaherty, Roisin
AU  - O'Flaherty R
AD  - NIBRT GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research 
      and Training, Fosters Avenue, Mount Merrion, Blackrock, Co., Dublin, Ireland.
FAU - Karlsson, Niclas G
AU  - Karlsson NG
AD  - Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, 
      Sahlgrenska Academy, University of Gothenburg, Box 440, 405 30, Gothenburg, 
      Sweden.
FAU - Rudd, Pauline M
AU  - Rudd PM
AD  - NIBRT GlycoScience Group, NIBRT-The National Institute for Bioprocessing Research 
      and Training, Fosters Avenue, Mount Merrion, Blackrock, Co., Dublin, Ireland. 
      pauline.rudd@nibrt.ie.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Glycoproteins)
RN  - 0 (Polysaccharides)
SB  - IM
MH  - Chromatography, High Pressure Liquid/instrumentation/*methods
MH  - Equipment Design
MH  - Glycomics/instrumentation/*methods
MH  - Glycoproteins/blood/*chemistry/isolation & purification
MH  - Glycosylation
MH  - High-Throughput Screening Assays/instrumentation/methods
MH  - Humans
MH  - Plasma/chemistry
MH  - Polysaccharides/*analysis/blood/isolation & purification
MH  - Protein Denaturation
MH  - Software
MH  - Solid Phase Extraction/instrumentation/methods
OTO - NOTNLM
OT  - Automation
OT  - Glycan analysis
OT  - High-throughput
OT  - N-linked glycosylation
OT  - Plasma
OT  - Robotics
OT  - Ultrahigh-performance liquid chromatography
EDAT- 2016/10/16 06:00
MHDA- 2018/01/18 06:00
CRDT- 2016/10/16 06:00
PHST- 2016/10/16 06:00 [entrez]
PHST- 2016/10/16 06:00 [pubmed]
PHST- 2018/01/18 06:00 [medline]
AID - 10.1007/978-1-4939-6493-2_8 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2017;1503:97-108. doi: 10.1007/978-1-4939-6493-2_8.