COMPARISON OF DENATURING AGENTS USED IN N-GLYCAN RELEASE METHODS FOR QUANTITATIVE GLYCOMIC ANALYSIS
Proje Ekibi (1)
| Proje Türü | Yükseköğretim Kurumları Tarafından Destekli Bilimsel Araştırma Projesi (Yükseköğretim Kurumları tarafından destekli bilimsel araştırma projesi) |
| Proje No | - |
| Proje Sahibi Kurum | Karabük Üniversitesi |
| Proje Durumu | Tamamlandı |
| Başlangıç Tarihi | 27-05-2020 |
| Bitiş Tarihi | 21-12-2022 |
| Proje Konusu | Glycosylation is an essential post-translational modification observed in the living proteome. Complex oligosaccharides called glycans are attached to proteins in this process and, the functions of proteins are changed dramatically. In addition, glycosylation profiles can change in commonly observed diseases such as cancer. Identifying these changes is crucial in discovering new biomarkers for the early detection of cancer and the development of new targeted drugs. Therefore, N-glycan profiling of glycoproteins extracted from various biological samples is crucial. One of the main steps of N-glycan analysis is to remove N-glycans from glycoproteins by specific enzymes. The study aims to compare detergent combinations used in N-glycan release methods. In the study, human plasma was used to test the release methods of N-glycans containing different detergent combinations. The released N-glycans were labeled with the procainamide tag and purified using cellulose-containing solid-phase extraction cartridges. Analyses were performed with HPLC-HILIC-FLD (high-performance liquid chromatography-hydrophilic interaction liquid chromatography equipped with fluorescence detection), which is the golden standard method for profiling N-glycans. The results showed that the, sodium dodecyl sulfate + sodium deoxycholate detergent combination provided the most efficiency in the N-glycan analysis. It was found that the total and average signal areas and intensities of the detected N-glycans were reduced when SDS and SDC detergents combined with DTT. In addition, deglycosylation of glycoproteins with various denaturing agents resulted in different human plasma N-glycosylation profiles. |
