Contacts:
Supervisor(s):
Prof. Fabio Di Francesco, Dr. Tommaso Lomonaco
Title of the PhD project:
Mass spectrometry-based assay for the determination of natriuretic peptides in saliva and plasma samples.
Abstract of the PhD project:
Cardiac natriuretic peptides (NPs), i.e., ANP, BNP, and CNP, are hormones initially synthesized from cardiomyocytes, as prohormones. Once they are released from the heart, prohormones are split into two fragments: a longer N-terminal fragment (NT-proBNP and NT-proANP) and a shorter C-terminal fragment (BNP and ANP). In pathological conditions as heart failure (HF), heart induces the stretching of myocytes due to pressure overload, leading to the release of NPs in blood. Thus, NPs plasma levels increase progressively with the severity of HF and represent the main biomarkers for prognosis and diagnosis of HF. NPs are normally determined in blood using commercial immunoassay kits. This approach is affected by cross-reactions due to the presence of several NP forms showing homologous structures, as those with oxidized methionine. My project aims to the development of a reliable and innovative analytical workflow based on targeted top-down proteomics and UHPLC-MS/MS analysis for the determination of NPs in saliva and plasma samples. The ability to rapidly quantify multiple molecular species with high specificity and accuracy makes mass spectrometry suitable for determining the different NPs forms. Ultrafiltration, fractional protein precipitation, and micro-extraction by packed sorbent (MEPS) will be tested as sample treatment procedures. Finally, the optimized analytical protocol will be employed for the longitudinal monitoring of patients suffering from HF, and for a systematic comparison between the analytical performances of the proposed method and immunoassays used in the clinical routine for the quantification of NPs.
Publications:
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Oral communications at conferences:
• A. Lenzi, D. Biagini, S. Ghimenti, F. Vivaldi, A. Bonini, P. Salvo, R. Fuoco, F. Di Francesco, T. Lomonaco “Determination of salivary short chain fatty acids and hydroxy acids in heart failure patients by in-situ derivatization and HiSorb-probe sorptive extraction coupled to TD-GC-MS/MS”, 9th MS J Day, Oral presentation, Online Edition (June 24, 2021).
• A. Lenzi, D. Biagini, S. Ghimenti, F. Vivaldi, A. Bonini, P. Salvo, R. Fuoco, F. Di Francesco, T. Lomonaco “Determination of salivary short chain fatty acids and hydroxy acids in heart failure patients by in-situ derivatization and HiSorb-probe sorptive extraction coupled to TD-GC-MS/MS”, XXVII Congresso della Società Chimica Italiana 2021, Oral presentation, Online Edition (September 14 - 23, 2021).
• A. Lenzi, M. De Cristofaro, D.Biagini, S. Ghimenti, F. Di Francesco, T. Lomonaco “Mass spectrometry-based assay for the determination of natriuretic peptides in plasma samples”, XXIX Congresso della Divisione di Chimica Analitica 2022, Oral presentation, Milazzo (ME) (September 11 - 25, 2022).
• A. Lenzi, M. De Cristofaro, D.Biagini, S. Ghimenti, F. Di Francesco, T. Lomonaco “MS-based assay for the determination of natriuretic peptides in plasma and saliva samples”, Autumn Meeting for Young Chemists in Biomedical Sciences 2022, Slide&Talk, Napoli (October 17 - 19, 2022).
• A. Lenzi, M. De Cristofaro, D.Biagini, S. Ghimenti, F. Di Francesco, T. Lomonaco “Development of a liquid chromatography-tandem mass spectrometry method to quantify an array of natriuretic peptides in human plasma”, Merck Young Chemists' Symposium 2022, Oral Presentation, Rimini (November 21 - 23, 2022).
Poster communications at conferences:
• A. Lenzi, D. Biagini, S. Ghimenti, F. Vivaldi, A. Bonini, P. Salvo, R. Fuoco, F. Di Francesco, T. Lomonaco “Determination of salivary short chain fatty acids and hydroxy acids in heart failure patients by in-situ derivatization and HiSorb-probe sorptive extraction coupled to TD-GC-MS/MS”, Chemistry for the Future 2021, Online Edition (June 30 - July 2, 2021).
• A. Lenzi, M. De Cristofaro, D.Biagini, S. Ghimenti, F. Di Francesco, T. Lomonaco “Development of a targeted top-down proteomic approach for determining natriuretic peptides in biological fluids”, Chemistry for the Future 2022, Pisa, (June 29 - July 1, 2022).
Other achievements:
Best Poster Award: Chemistry for the Future 2022, Pisa, June 29-July 1, 2022
