Here, we prepared a magnetic nanocomposite system composed of a cluster of magnetite nanoparticles coated with silica shell (MSNPs) with an average diameter of 140 ± 20 nm and conjugated with CD9 antibody (AntiCD9) using different strategies including adsorption or chemical conjugation of antibody molecules to either aminated MSNPs (AMSNPs) or carboxylated MSNPs (CMSNPs). Then, MSNPs were employed to isolate exosomes from ultracentrifuge-enriched solution, PC3 cell-culture medium, or exosome-spiked simulated plasma samples. Quantitative tests using nanoparticle-tracking analysis confirmed antibody-covalently conjugated MSNPs, i.e. the AntiCD9-AMSNPs and AntiCD9-CMSNPs enabled >90% recovery of exosomes. Additionally, the exosomes isolated with AntiCD9-CMSNPs showed higher recovery efficiency compared to the AntiCD9-AMSNPs. For both nanoadsorbents, lower protein impurities amounts were obtained as compared to that of exosomes isolated by ultracentrifugation and Exocib kit. The mean diameter assessment of the isolated exosomes indicates that particles isolated by using AntiCD9-AMSNPs and AntiCD9-CMSNPs have smaller sizes (136 ± 2.64 nm and 113 ± 11.53 nm, respectively) than those obtained by UC-enriched exosomes (140.9 ± 1.6 nm) and Exocib kit (167 ± 10.53 nm). Such promising results obtained in the isolation of exosomes recommend magnetic nanocomposite as an efficient tool for the simple and fast isolation of exosomes for diagnosis applications.
Keywords: antibody; exosome; immobilization; isolation; magnetic nanoadsorbents.
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