Magnetic Bead-based Exosome Extraction: A Technique to Isolate Specific Exosomes from Biofluids

Abstract

Source: Tu, M. et al. Detection of Exosomal Biomarkers by Electric Field-induced Release and Measurement (EFIRM). J. Vis. Exp. (2015)

In this video, we demonstrate a procedure to isolate exosomes from target biofluids using magnetic microbeads. The procedure enables the rapid capture of exosomes, which can be used for downstream analysis.

Protocol

1. Magnetic Bead-based Exosome Extraction

1. Pipette a well-mixed solution of 5 µL of Streptavidin-coated magnetic microparticles into 495 µL of phosphate-buffered saline (PBS) buffer in a microcentrifuge tube to resuspend the beads. Wash and resuspend the beads with 500 µL of PBS three times using a magnetic rack. The rack is an array of magnets on the side of a housing unit that can hold the sample microcentrifuge tubes.
   1. For each wash, first, let the tubes sit on the rack for 1 min and then use a pipette tip to carefully remove the supernatant buffer without disturbing the beads.
   2. Place the tubes on a re....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Biotinylated anti-human CD63 Antibody	Ancell, USA	215-030
Dynabeads MyOne Streptavidin T1	Invitrogen, USA	65601
Neodynium Magnetics (1/10" dia. x 1/32" thick)	K&J Magnetics, USA	DH101
Ultrapure Distilled Water	Life Technologies, USA	10977-023
Anti-Fluorescein-POD, Fab fragments	Roche, Germany	11426346910
Phosphate Buffered Saline Solution	Life Technologies, USA	10010023
Casein/PBS	Fisher Scientific, USA	37532