Development of efficient on-bead protein elution process coupled to ultra-high performance liquid chromatography-tandem mass spectrometry to determine immunoglobulin G subclass and glycosylation for discovery of bio-signatures in pancreatic disease
Journal
Journal of chromatography. A
Journal Volume
1621
Date Issued
2020-06-21
Author(s)
Shiao, Jing-Ya
Chen, Michael X
Liu, Li-Wei
Wang, Xiang-Yu
Tsai, Yun-Jung
Kuo, Tai-Chih
Tsai, I-Lin
Abstract
Type 1 autoimmune pancreatitis (AIP) is a kind of IgG4-related disease in which higher IgG4 and total IgG levels have been found in patient serum. Due to the similar imaging features and laboratory parameters between AIP and pancreatic ductal adenocarcinoma (PDAC), a differential diagnosis is still challenging. Since IgG profiles can be potential bio-signatures for disease, we developed and validated a method which coupled on-bead enzymatic protein elution process to an efficient UHPLC-MS/MS method to determine IgG subclass and glycosylation. A stable-isotope labeled IgG was incorporated as internal standard to achieve accurate quantification. For calibration curves, the correlation coefficients for total IgG and the four IgG subclasses were higher than 0.995. Intraday (n = 5) and interday (n = 3) precisions of the peak area ratios of LLOQ, low, medium, and high QC samples were all less than 6.6% relative standard deviation (% RSD), and the accuracies were between 93.5 and 114.9%. Calibration curves, precision, and accuracy were also evaluated for 26 IgG glycopeptides. The method was applied to samples from healthy controls and patients with AIP and PDAC. Distinct IgG patterns were discovered among the groups, and 7 glycopeptides showed high potential in differentiating AIP and PDAC. The results demonstrated that the developed method is suitable for multi-feature analysis of human IgG, and the discovered IgG profiles can be used as bio-signatures for AIP and PDAC.
Subjects
Autoimmune pancreatitis; Fc N-glycan; Igg; LC–MS/MS; Pancreatic cancer; Serum
SDGs
Other Subjects
Calibration; Glycosylation; High performance liquid chromatography; Mass spectrometry; Peptides; Accurate quantifications; Calibration curves; Correlation coefficient; Differential diagnosis; Ductal adenocarcinomas; Relative standard deviations; Tandem mass spectrometry; Ultra-high performance liquid chromatographies; Diagnosis; glycopeptide; immunoglobulin G; glycopeptide; immunoglobulin G; Article; autoimmune pancreatitis; clinical article; controlled study; correlation analysis; elution; glycosylation; human; isotope labeling; limit of quantitation; measurement accuracy; pancreas adenocarcinoma; pancreas disease; priority journal; protein processing; signal noise ratio; tandem mass spectrometry; ultra performance liquid chromatography; validation study; autoimmune pancreatitis; blood; classification; differential diagnosis; glycosylation; high performance liquid chromatography; immunology; metabolism; pancreas carcinoma; pancreas tumor; procedures; tandem mass spectrometry; Autoimmune Pancreatitis; Carcinoma, Pancreatic Ductal; Chromatography, High Pressure Liquid; Diagnosis, Differential; Glycopeptides; Glycosylation; Humans; Immunoglobulin G; Pancreatic Neoplasms; Tandem Mass Spectrometry
Publisher
ELSEVIER
Type
journal article