title: Point-of-care detection of extracellular vesicles: Sensitivity optimization and multiple-target detection.
abstract: Extracellular vesicles (EVs) are membrane-bound nanovesicles delivered by different cellular lineages under physiological and pathological conditions. Although these vesicles have shown relevance as biomarkers for a number of diseases, their isolation and detection still has several technical drawbacks, mainly related with problems of sensitivity and time-consumed. Here, we reported a rapid and multiple-targeted lateral flow immunoassay (LFIA) system for the detection of EVs isolated from human plasma. A range of different labels (colloidal gold, carbon black and magnetic nanoparticles) was compared as detection probe in LFIA, being gold nanoparticles that showed better results. Using this platform, we demonstrated that improvements may be carried out by incorporating additional capture lines with different antibodies. The device exhibited a limit of detection (LOD) of 3.4×106EVs/µL when anti-CD81 and anti-CD9 were selected as capture antibodies in a multiple-targeted format, and anti-CD63 labeled with gold nanoparticles was used as detection probe. This LFIA, coupled to EVs isolation kits, could become a rapid and useful tool for the point-of-care detection of EVs, with a total analysis time of two hours.
authors: Departamento de Química Física y Analítica, Universidad de Oviedo, Julián Clavería 8, Oviedo, 33006 Spain.
authors: Myriam
authors: M
authors: Oliveira-Rodríguez
authors: Departamento de Ingeniería Química y Tecnología del Medio Ambiente, Universidad de Oviedo, Julián Clavería 8, Oviedo, 33006 Spain.
authors: Esther
authors: E
authors: Serrano-Pertierra
authors: Departamento de Química Física y Analítica, Universidad de Oviedo, Julián Clavería 8, Oviedo, 33006 Spain.
authors: Agustín Costa
authors: AC
authors: García
authors: Unidad de Investigación, Hospital St Cristina, Instituto de Investigación Sanitaria Princesa (IS-IP), Madrid, Spain.
authors: Soraya
authors: S
authors: López-Martín
authors: Unidad de Investigación, Hospital St Cristina, Instituto de Investigación Sanitaria Princesa (IS-IP), Madrid, Spain; Departamento de Biología Molecular, UAM/CBM-SO, Madrid, Spain.
authors: María
authors: M
authors: Yañez-Mo
authors: Servicio de Neurología, Hospital Universitario Central de Asturias, Oviedo, Spain.
authors: Eva
authors: E
authors: Cernuda-Morollón
authors: Departamento de Química Física y Analítica, Universidad de Oviedo, Julián Clavería 8, Oviedo, 33006 Spain. Electronic address: cblanco@uniovi.es.
authors: M C
authors: MC
authors: Blanco-López
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015374:Biosensing Techniques; D004867:Equipment Design; D000067128:Extracellular Vesicles / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D017979:Gold Colloid / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation; D018796:Immunoconjugates / Q000737:chemistry; D057230:Limit of Detection; D058185:Magnetite Nanoparticles / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D019095:Point-of-Care Systems; D060225:Tetraspanin 28 / Q000032:analysis*; D060245:Tetraspanin 29 / Q000032:analysis*
chemical_list: D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D017979:Gold Colloid; D018796:Immunoconjugates; D058185:Magnetite Nanoparticles; D060225:Tetraspanin 28; D060245:Tetraspanin 29; D006046:Gold
keywords: Extracellular vesicles; Lateral flow immunoassay; Multiple-targeted detection; Nanoparticles; Point-of-care