Introduction

The invention consists of an innovative procedure, easily applicable, for the rapid ed efficient isolation of extracellular vesicles (EVs). The method allows the purification of intact nanoparticles in physiological solution and the integration to ultrasensitive and homogeneous systems for detecting disease biomarkers (proteins and nucleic acids).

Technical features

Extracellular vesicles (EVs) are spherical particles (nano- and sub-micrometric) massively secreted from the cells in different biological fluids, such as blood, urine, saliva, etc. The invention consists of a method for capturing these particles based on the net negative charge that characterizes them. The main novelty lies in the selective elution of EVs from the stationary phase, which can be obtained in a physiological solution that preserves EV integrity and stability. In addition, the possibility to obtain single particles in solution allows to analyze EVs with ultrasensitive technologies, like amplified luminescent proximity homogeneous assay (alpha) and droplet digital PCR (ddPCR), for detecting proteins or nucleic acids, achieving unprecedented sensitivity from liquid biopsies.

Possible Applications

• Isolation of nanoparticles for research purposes;
• Analysis of nanoparticles for molecular diagnosis of cancer or neurodegenerative diseases;
• Use of nanoparticles for therapeutic purposes as drug-loaded or genome editing vectors.

Advantages

• Label free approach (independent on antibodies);
• Fast procedure (EVs in less than 1 hr);
• Efficient/scalable system (up to 98% recovery of EVs from different volumes of biofluid);
• Reproducible and versatile system (it can be combined with traditional or next-generation analytical devices).