BIOMARKER ENRICHMENT TOOLS UTILISING THE NANOPARTICLE BIOMOLECULE CORONA

Abstract

Mass spectrometry (MS) is the predominant technology to discover protein biomarkers for diseases. Despite the escalated investment on cutting-edge instruments and the availability of manifold MS-based quantitation techniques, only a few plasma biomarkers are clinically used. The highly dynamic environment of plasma along with the overwhelming signal of the most abundant proteins restrict the identification of low abundant and low molecular weight (MW) species, most likely to be biomarker candidates. With the current state-of-the-art solutions failing to tackle these limitations, the latest technological breakthroughs in nanomedicine have posed pronounced hope for plasma biomarker discovery. Recent studies have shown that nanoparticles (NPs) enrich complex biofluids with low abundant and low MW disease-specific biomolecules, previously undetectable with conventional MS-proteomics, following their spontaneous interaction with plasma proteins referred to as the ‘biomolecule corona’. In this thesis, we aim to further exploit this plasma enrichment nanoplatform for previously unexplored clinical scenarios. Initially, we develop a robust workflow for the in-depth MS analysis of the human plasma-derived liposome-biomolecule corona. Our data reveal that the proposed protocol generates a purified sample with high MS reproducibility, and signify the great advantage of using the biomolecule corona over plasma alone for proteomic analysis. Furthermore, the liposome-corona is examined as a scavenging tool for the discovery of novel protein biomarkers in two previously unexplored disease cases: Parkinson’s Disease (PD) and sepsis. Interestingly, multiple disease-specific protein signatures are unveiled in plasma for both cases following the proteomic comparison of ‘healthy’ and ‘diseased’ coronas. Lastly, we provide the first evidence of the presence of a broad range of bioactive lipids onto liposomes, suggesting that the biomolecule corona enriches the lipidomic fingerprints of plasma. In summary, this work reinforces the novelty of the NP-biomolecule corona as a proteomics enrichment tool to harvest disease-specific biomarkers, reveals its rich content in plasma lipids and highlights the future use of combinatorial omics to globally characterise it.

Date of Award	31 Dec 2022
Original language	English
Awarding Institution	The University of Manchester
Supervisor	Richard Unwin (Supervisor), David Knight (Supervisor), Kostas Kostarelos (Supervisor) & Marilena Hadjidemetriou (Supervisor)