NEXUS · automated in-line separatioN and dEtection of eXtracellular vesicles for liqUid biopsy applicationS

The NEXUS project will develop an easily customizable integrated platform to enable Extracellular Vesicles (EVs) separation, purification and multivariate characterization from complex biofluids. EVs enclose a heterogeneous group of cell-released membranous vesicles, abundantly present in body fluids from which they can be extracted in a non-invasive manner. Their surface markers and cargo (proteins, nucleic acids, glycans, lipids) represent an ideal source of diagnostic, prognostic and efficacy biomarkers, providing real-time information on tissue homeostasis alterations. However, their detection and molecular profiling is technically challenging due to their physical characteristics and huge heterogeneity. The previous FET project INDEX has validated at lab scale (TRL4) an integration of a selective sequential isolation and in situ enrichment of small EVs (sEVs) from plasma, with the multiparameter analysis regarding vesicle size, number and immunophenotype. NEXUS aims at proceeding towards industrialisation by delivering a manufacturable, full-fledged analytical instrument, characterised by an appealing ?sample in ? result out? design and featuring a real time in liquid measurement of scalable sample volumes. NEXUS breakthrough technology will be achieved through integration of: i) innovative chemistry for reversible sEVs immunocapture with unique Capture-Release-Re-capture features; ii) microfluidic extraction and pre-concentration of cancer sEVs; iii) DNA barcoded customizable chips for single sEV capture and analysis. Fully automated Interferometric Reflectance Imaging Sensor providing rapid, sensitive and multiplex profiling of cancer proteins, displayed on sEVs without the interference of soluble proteins and confounding particles (other sEVs and lipoproteins), will be validated in the relevant industrial and clinical environment and in the context of a Liquid Biopsy assay for Prostate Cancer Stratification, Prognosis and Monitoring and Prognosis (TRL6).