CESI-MS for analysis of disease induced protein modifications
Jenny Van Eyk
Professor of Medicine, Advanced Clinical Biosystems Research Institute, Barbara Streisand Women's Heart Center, Heart Institute
Heart disease is the number one cause of mortality worldwide, and risk stratification for developing heart failure (HF) and/or to monitoring HF progression is needed. The two primary biomarkers used in heart disease, cardiac troponin I (cTnI) and B-type Natriuretic Peptide (BNP), both have disease-induced modifications that we are studying in an effort to produce markers with increased disease state specificity.
The goal is to analytically define BNP variants in patient samples to provide a more comprehensive clinical assessment. The challenge is that cleaved BNP rapidly degrades once released into the blood. Proteoforms exist transiently and at low concentrations in plasma. Using capillary electrophoresis coupled to mass spectrometry (CESI-MS), we are able to analyze multiple BNP cleavage products in plasma without resorting to expensive reagents or complex instrumentation. We have developed assays that are able to provide quantitative information about the proteolysis rate of BNP in blood. Our method includes minimal sample processing that can be completed in under ten minutes. We ultimately aim to profile BNP cleavage in a plasma sample, including sample processing and analysis, in less than one hour.
Key Learning Objective:
- How capturing post-translational modifications (PTMs) that occur due to pathological processes provide us with opportunities to identify biomarkers that are specific for a particular disease state.
- Learn about the quantitative assay developed for analyzing BNP and its proteolytic variants, using minimum sample processing, and in less than 10 minutes processing time.
- How CESI-MS is used to analyze multiple BNP cleavage products in plasma without resorting to expensive reagents or complex instrumentation.