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A Microfluidic ExoNANO for Efficient Isolation and Chemiluminescence Quantitative Immunoassay of Exosomes
30 Pages Posted: 10 Dec 2018
More...Abstract
Background: Although exosomes are potential diagnostic and therapeutic biomarkers for diverse diseases, efficient and sensitive technologies for exosomes isolation and detection are still lacking.
Methods: In this study, we construct a microfluidic system (ExoNANO) for exosomes separation using CD63 antibody conjugated nano-sized superparamagnetic iron oxide particles (SIOP) and microfluidic isolation column. Further, a chemiluminescence immunoassay (CLIA) strategy was established for exosomal protein detection.
Findings: Results show that microfluidic ExoNANO reveals high isolation efficiency (94%) and allows quantitative analysis of exosomes in the range of 2.92×105 to 2.80×108 particles/μL with a limit of detection (LOD) of 2.63×105 particles/μL from as little as 1μL serum sample. Using microfluidic ExoNANO, we quantified exosomes in serum, cell culture media (CM), saliva, ascitic fluid and cerebrospinal fluid (CSF) samples.
Interpretation: We developed microfluidic ExoNANO, which could pave the way for routine, rapid isolation and quantitative analysis of exosomes in clinical application.
Funding Statement: This study was supported by grant from the National Natural Science Foundation of China (Grant nos. 81271917, 81802571), Natural Science Foundation of Zhejiang province (Grant nos. LY14H200002, LY15H200002 and LY16H160023).
Declaration of Interests: The authors declare: "There does not exist any conficts of interest."
Ethics Approval Statement: Serum samples of healthy individuals and cerebrospinal fluid, ascitic fluid, and saliva samples were obtained from the Second Affiliated Hospital of Zhejiang University School of Medicine. Research protocols were approved by the Ethical Committees for Clinical Research of supporting institutions, and all patients provided written informed consent for the use of their samples for experimentation.
Keywords: microfluidic, exosomes, superparamagnetic iron oxide particles, chemiluminescence immunoassay
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