Magnetic Separation of Cell-Secreted Vesicles with Tailored Magnetic Particles and Downstream Applications

Mireia Bernuz, Arnau Pallarès-Rusiñol, Rosanna Rossi, Carolina Fernández-Senac, Mercè Martí, María Isabel Pividori*

*Autor corresponent d’aquest treball

Producció científica: Capítol de llibreCapítolRecercaAvaluat per experts

Resum

The analysis of the receptors on the surface of the cell-secreted vesicles provides valuable information of the cell signature and may also offer diagnosis and/or prognosis of a wide range of diseases, including cancer. Due to their low concentration, conventional procedures for extracellular vesicle (EV) detection usually require relatively large sample volumes, involving preliminary purification or preconcentration steps from complex specimens. Here, we describe the separation and preconcentration in magnetic particles of extracellular vesicles obtained from cell culture supernatants from MCF7, MDA-MB-231, and SKBR3 breast cancer cell lines, human fetal osteoblastic cell line (hFOB), and human neuroblastoma SH-SY5Y cell line, as well as exosomes from human serum. The first approach involves the covalent immobilization for the exosomes directly on micro (4.5 μm)-sized magnetic particles. The second approach is based on tailored magnetic particles modified with antibodies for further immunomagnetic separation of the exosomes. In these instances, micro (4.5 μm)-sized magnetic particles are modified with different commercial antibodies against selected receptors, including the general tetraspanins CD9, CD63, and CD81 and the specific receptors (CD24, CD44, CD54, CD326, CD340, and CD171). The magnetic separation can be easily coupled with downstream characterization and quantification methods, including molecular biology techniques such as immunoassays, confocal microscopy, or flow cytometry.

Idioma originalEnglish
Títol de la publicacióMethods in Molecular Biology
Pàgines257-276
Nombre de pàgines20
ISBN (electrònic)978-1-0716-3203-1
DOIs
Estat de la publicacióPublicada - 2023

Sèrie de publicacions

NomMethods in Molecular Biology
Volum2668
ISSN (imprès)1064-3745
ISSN (electrònic)1940-6029

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