Abstract
Quantitative detection of exosomes in bio-fluids is a challenging task in a dynamic research field. The absence of a well-established reference material (RM) for method development and inter-comparison studies could be potentially overcome with artificial exosomes: lab-produced biomimetic particles with morphological and functional properties close to natural exosomes. This work presents the design, development and functional characteristics of fully artificial exosomes based on tetraspanin extracellular loops-coated niosomes, produced by bio-nanotechnology methods based on supra-molecular chemistry and recombinant protein technology. Mono- and double-functionalized particles with CD9/CD63 tetraspanins have been developed and characterized from a morphological and functional point of view. Produced bio-particles showed close similarities with natural entities in terms of physical properties. Their utility for bioanalysis is demonstrated by their detection and molecular-type discrimination by enzyme-linked immunosorbent assays (ELISAs), one of the most frequent bio-analytical method found in routine and research labs. The basic material based on streptavidin-coated niosomes allows the surface functionalization with any biotinylated protein or peptide, introducing versatility. Although promising results have been reported, further optimizations and deeper characterization will help this innovative biomaterial become a robust RM for validation and development of diagnostic tools for exosomes determination.
Highlights
Extracellular vesicles have emerged as a novel mechanism of intercellular communication over the last years, playing an important role in both biological and pathological processes [1]
We described for the first time double functionalized particles (CD9 and CD63), and their functionality was tested in an enzyme-linked immunosorbent assays (ELISAs) assay
The strategies for the development of artificial exosomes have been reviewed in a previous publication of our group [22]
Summary
Extracellular vesicles have emerged as a novel mechanism of intercellular communication over the last years, playing an important role in both biological and pathological processes [1]. Exosomes are vesicular subcellular particles with an average size around 100–150 nm in diameter, characterized by a particular protein profile which offers valuable information. This information can be about the cell from which they are released, their target cell population, about the health status of the organism/cell, and their possible (patho)physiological roles [3,4]. Their possibilities as biomarkers for diagnosis [5] and treatment-response monitoring [6] promote their determination in biological fluids and cell culture media as a routine practice in cell biology laboratories, and in the clinical research. Several strategies for EVs isolation, detection and quantification have been developed, as reviewed elsewhere [8,9,10]
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