Detection and Quantitation of β-2-Microglobulin Glycosylated End Products in Human Serum by Matrix-Assisted Laser Desorption/lonization Mass Spectrometry

摘要

β-2-Microglobulin (β_2M) is a major protein component found in the amyloid deposits of dialysis-related amyloi-dosis (DRA) patients. Evidence has been shown that the advanced glycosylated end-products (AGEs) of β_2M present in sera were related to DRA. We demonstrated mat matrix-assisted laser desorption/ionization mass spec-trometry (MALDI-MS) is a useful tool to investigate the nature of glycosylanon of β_2M and detection of β_2M or β_2M-AGEs in human serum. The high-mass end of β_2M-AGE distribution was found to extend to me neighborhood of 12 868 Da, corresponding to condensations with seven glucose molecules. We also have shown that both β_2M and β_2M-AGEs can be detected at low picomole levels directly in bovine serum. Based on these findings, the sera of DRA patients were studied to determine whether β_2M-AGEs can be detected by MALDI-MS. In an attempt to investigate the possibility of quantitation with MALDI, human sera samples with different concentrations of β_2M-AGE were examined. We were able to correlate the concentration of β_2M-AGE with the number of detected AGE products, pointing to the feasibility of MALDI as a quantitative tool.