Comparative Analysis and Validation of Different Steps in Glycomics Studies (CROSBI ID 64065)
Prilog u knjizi | izvorni znanstveni rad | međunarodna recenzija
Podaci o odgovornosti
Trbojević-Akmačić, Irena ; Ugrina, Ivo ; Lauc, Gordan
engleski
Comparative Analysis and Validation of Different Steps in Glycomics Studies
Large-scale glycomics studies enable identification of aberrant glycosylation patterns in disease and provide information about functional relevance of individual glycans through genome-wide association studies. Developed high-throughput methodologies have to be sensitive, robust, and stable during long periods of time (few months) to be able to reliably detect small biological variations in glycosylation. Here, we describe a simple, robust, and affordable protocol for immunoglobulin G N-glycan analysis by hydrophilic interaction liquid chromatography–ultra-performance liquid chromatography (HILIC-UPLC), as well as useful strategies for method optimization: Plackett–Burman screening design and analysis of source of variation. We put our focus on experimental design for high-throughput glycan analysis, critical steps in sample preparation procedure for obtaining high-quality data, and propose a validation protocol relevant for high-throughput methods in terms of their long-term robustness and ability to detect biologically relevant changes in glycosylation. The quality of the procedure was assessed by employing appropriate experimental designs and subsequent statistical techniques.
Glycans ; High throughput ; Glycome analysis ; HILIC ; UPLC ; Method validation
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Podaci o prilogu
37-55.
objavljeno
10.1016/bs.mie.2016.09.027
Podaci o knjizi
Proteomics in Biology, Part B
Shukla, Arun K.
Cambridge (MA): Elsevier
2017.
978-0-12-809743-4
0076-6879