Applications of Mass Spectrometry Proteomic Methods to Immunoglobulins in the Clinical Laboratory DOI
David Murray, Maria Alice V. Willrich

Clinical Chemistry, Journal Year: 2024, Volume and Issue: 70(12), P. 1422 - 1435

Published: Nov. 27, 2024

Abstract Background Immunoglobulin (Ig) measurements in the clinical laboratory have been traditionally performed by nephelometry, turbidimetry, electrophoresis, and ELISA assays. Mass spectrometry (MS) potential to provide deeper insights on nature of these markers. Content Different approaches—top-down, middle-down, or bottom-up—have described for measuring specific Igs endogenous monoclonal immunoglobulins (M-proteins) exogenous therapeutic antibody therapies (t-mAbs). Challenges arise distinguishing Ig interest from polyclonal background. MS is emerging as a practical method quantitative analysis information about structural clonal features that are not easily determined current methods. This review discusses clinically implemented examples, including isotyping quantification M-proteins quantitation t-mAbs within background, examples how can enhance our detection characterization Igs. Summary available proteomic tests highlights both analytical nonanalytical challenges implementation. Given new insight into methods, it hoped vendors, laboratorians, healthcare providers, payment systems work overcome advance care patients.

Language: Английский

B4GALT3 as a Key Glycosyltransferase Gene in Multiple Myeloma Progression: Insights from Bioinformatics, Machine Learning, and Experimental Validation DOI Creative Commons

Apeng Yang,

Mengying Ke,

Feng Lin

et al.

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 28, 2025

Abstract Background: Glycosylation abnormalities are critical in the progression of various cancers. However, their role onset and prognosis multiple myeloma (MM) remains underexplored. This study aims to identify glycosyltransferase (GT)-related biomarkers investigate underlying mechanisms MM. Methods: GT-related genes were extracted from MMRF-CoMMpass GSE57317 datasets. Potential identified using Cox regression Lasso analyses. A Glycosyltransferase-Related Prognostic Model (GTPM) was developed by evaluating 113 machine learning algorithm combinations. The expression B4GALT3, a key gene through this model, analyzed MM bone marrow samples immunohistochemistry, quantitative PCR, western blotting. Functional roles B4GALT3 cell behavior assessed knockdown experiments, its mechanism action investigated. Results: GTPM stratified patients into high- low-risk groups, with significantly better survival group (HR = 55.94, 95% CI 40.48–77.31, p \(&#x003C;\) 0.001). model achieved AUC values 0.98 0.99 for 1-year 3-year overall survival, outperforming existing signatures (including EMC92, UAMS70, UAMS17). elevated advanced stages (p $<$ 0.001) correlated poorer survival. Knockdown reduced proliferation, invasion , increased apoptosis. Mechanistic analyses revealed that modulates via Wnt/ \(\beta\) -catenin/GRP78 pathway, primarily regulating endoplasmic reticulum (ER) stress. Conclusions: novel predicting as influencing disease progression. Experimental evidence highlights B4GALT3's modulating ER stress Wnt/\(\beta\)-catenin pathways, positioning it potential prognostic biomarker therapeutic target

Language: Английский

Citations

0
Deciphering N-Glycosylation Dynamics of Serum Monoclonal Immunoglobulins in Multiple Myeloma via EThcD-sceHCD-MS/MS DOI
Huixian Li,

Wanhong Lu,

Jin Qian

et al.

Journal of Proteome Research, Journal Year: 2025, Volume and Issue: unknown

Published: April 9, 2025

Serum glycoprotein glycosylation changes can indicate disease onset and progression. However, the site-specific N-glycosylation of monoclonal immunoglobulins (M-proteins) in multiple myeloma (MM) its clinical implications are unclear. In this study, we isolated pathogenic micromonoclonal IgA or IgG (approximately 2 μg) from IgA-MM patients (n = 22) IgG-MM 30), normal polyclonal healthy controls (HCs) 16). Using EThcD-sceHCD-MS/MS, dynamics serum M-proteins MM were determined. Compared with IgA1 HCs, had higher fucosylation (58.1% vs 32.1%, p < 0.001), sialylation (68.0% 50.8%, 0.011), mannosylation (1.5% 0.3%, 0.001). While, IgG1 (97.8% 95.3%, addition, specific N-glycan abundances correlated features: for IgA1, HexNAc5Hex5Fuc1NeuAc1 was associated hypocomplementemia; IgG1, HexNAc4Hex3Fuc1 albumin level (r -0.363, 0.049) estimated glomerular filtration rate -0.433, 0.017); HexNAc4Hex5 therapeutic prognosis. conclusion, their isotypes HCs have distinct profiles, N-glycans characteristics

Language: Английский

Citations

0
Applications of Mass Spectrometry Proteomic Methods to Immunoglobulins in the Clinical Laboratory DOI
David Murray, Maria Alice V. Willrich

Clinical Chemistry, Journal Year: 2024, Volume and Issue: 70(12), P. 1422 - 1435

Published: Nov. 27, 2024

Abstract Background Immunoglobulin (Ig) measurements in the clinical laboratory have been traditionally performed by nephelometry, turbidimetry, electrophoresis, and ELISA assays. Mass spectrometry (MS) potential to provide deeper insights on nature of these markers. Content Different approaches—top-down, middle-down, or bottom-up—have described for measuring specific Igs endogenous monoclonal immunoglobulins (M-proteins) exogenous therapeutic antibody therapies (t-mAbs). Challenges arise distinguishing Ig interest from polyclonal background. MS is emerging as a practical method quantitative analysis information about structural clonal features that are not easily determined current methods. This review discusses clinically implemented examples, including isotyping quantification M-proteins quantitation t-mAbs within background, examples how can enhance our detection characterization Igs. Summary available proteomic tests highlights both analytical nonanalytical challenges implementation. Given new insight into methods, it hoped vendors, laboratorians, healthcare providers, payment systems work overcome advance care patients.

Language: Английский

Citations

0