Akademik Veri Yönetim Sistemi
International Conference of Biochemists and Molecular Biologists in Bosnia and Herzegovina, Sarajevo, Bosna-Hersek, 18 - 20 Mayıs 2023, ss.63, (Özet Bildiri)
Background: MS is a chronic inflammatory disease that causes demyelination throughout the central nervous system. The detection of oligoclonal bands (OCBs) in cerebrospinal fluid (CSF) using isoelectric focusing (IEF) is the gold standard method for MS diagnosis. However, its limited sensitivity necessitates the identification of accurate biomarkers, with IgG protein glycosylation motifs showing potential as such markers
Aim: The aim of this preliminary study was to isolate IgG proteins from serum and cerebrospinal fluid (CSF) samples using an IgG affinity matrix for subsequent glycomics analysis in MS patients.
Methods: In this study, IgG proteins from MS patients and controls serum and CSF samples were isolated and typed for OCBs using the IEF technique. IgG's were purified with the IgG affinity matrix and quantified with A280 measurement on a Nanodrop One. The IgG's heavy and light chains were separated with TCEP and analyzed for presence, structure, and purity using SDS-PAGE and MALDI-TOF analysis.
Results: IgG proteins were isolated and purified from serum and CSF using an IgG affinity matrix and typed for OCBs with IEF analysis. The purified IgG's heavy and light chains were separated and analyzed using SDS-PAGE and MALDI-TOF analysis. The light chains were found to have a size of 25 kDa, while the heavy chains had a size of 55 kDa in SDS-PAGE. The presence of lambda and kappa chains for both heavy and light chains was confirmed in the 11400-11800 m/z range using MALDI-TOF analysis. [M+2H]⁺² -[M+5H]⁺⁵ forms of IgG were also detected in the 10000-26000 m/z range.
Conclusion: In conclusion, the IgG affinity matrix shows promise for glycomics analysis in MS patients, with the potential to identify measurable biomarkers for early diagnosis and treatment. Further large-scale studies are needed to validate the use of IgG protein glycosylation motifs as MS diagnostic biomarkers.