Quantification of Specific Glycation Sites in Human Serum Albumin as Prospective Type 2 Diabetes Mellitus Biomarkers
| Autor: | Andrej Frolov, Sandro Spiller, Ralf Hoffmann |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Glycosylation Serum Albumin Human Plasma protein binding Biochemistry 03 medical and health sciences chemistry.chemical_compound Structural Biology Glycation medicine Humans Prospective Studies Solid phase extraction Binding Sites Type 2 Diabetes Mellitus General Medicine Human serum albumin Boronic Acids Blood proteins 030104 developmental biology Diabetes Mellitus Type 2 chemistry Biomarker (medicine) Peptides Biomarkers Protein Binding medicine.drug |
| Zdroj: | Protein & Peptide Letters. 24 |
| ISSN: | 0929-8665 |
| DOI: | 10.2174/0929866524666170202124120 |
| Popis: | Background Type 2 diabetes mellitus (T2DM) is the most common lifestyle disease affecting all countries. Due to its asymptomatic onset, it is often diagnosed after irreversible vascular complications have been initiated. Therefore, specific markers characteristic for very early disease stages and suitable for early diagnostics are required. Glycation of plasma proteins, such as human serum albumin (HSA), has been often suggested as marker. However, the total glycation degree of HSA does not provide sufficient information about short-term fluctuations of blood glucose concentrations due to the large number of glycation sites. Analysis of individual modification sites might be more informative, but methods for reliable quantifications are still missing. Objective The main objective of this study was to establish and qualify a method of analysis applicable to sensitive and precise quantification of glycations sites in plasma proteins. Methods Plasma samples obtained from diabetic patients and non-diseased individuals were separated from low-molecular weight compounds, digested with trypsin, enriched for glycated peptides by boronic acid affinity chromatography (BAC), desalted by solid phase extraction (SPE), and separated by RP-HPLC coupled online to ESI-QqQ-MS. Quantification relied on multiple reaction monitoring (MRM) of multiple glycation sites identified in plasma proteins using a stable isotope dilution approach or internal standardization. Results The data presented here suggests high selectivity and precision (relative standard deviations below 10%) of the overall approach appearing to be well suited for the identification of prospective biomarkers. Six glycated peptides corresponding to different glycation sites of HSA were present in plasma samples obtained from T2DM patients at significantly higher levels than in non-diabetic men matched for age. Additionally, each of the studied glycation site of HSA appeared to be affected at different degrees. Conclusion The presented approach enables the sensitive and robust quantification of prospective T2D biomarkers promising for clinical diagnostics. |
| Databáze: | OpenAIRE |
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