Naslov
Comparative Analysis and Validation of Different Steps in Glycomics Studies

Autori
Trbojević-Akmačić, Irena ; Ugrina, Ivo ; Lauc, Gordan

Vrsta, podvrsta i kategorija rada
Poglavlja u knjigama, znanstveni

Knjiga
Proteomics in Biology, Part B

Urednik/ci
Shukla, Arun K.

Izdavač
Elsevier

Grad
Cambridge (MA)

Godina
2017

Raspon stranica
37-55

ISBN
978-0-12-809743-4

ISSN
0076-6879

Ključne riječi
Glycans ; High throughput ; Glycome analysis ; HILIC ; UPLC ; Method validation

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija

POVEZANOST RADA