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A Microfluidic ExoNANO for Efficient Isolation and Chemiluminescence Quantitative Immunoassay of Exosomes

30 Pages Posted: 10 Dec 2018

See all articles by Yiyun Wang

Yiyun Wang

Zhejiang University

Zhenping Liu

Zhejiang University

Xuchu Wang

Zhejiang University

Yibei Dai

Zhejiang University

Xiaoying Li

Zhejiang University

Shifan Gao

Zhejiang University

Pan Yu

Zhejiang University

Qinyan Lin

Zhejiang University

Zihao Fan

Zhejiang University

Ying Ping

Zhejiang University

Danhua Wang

Zhejiang University

Xuedan Lin

Zhejiang University

Zejie Zheng

Zhejiang University

Weiwei Liu

Zhejiang University - Department of Laboratory Medicine

Zhihua Tao

Zhejiang University - Department of Clinical Laboratory

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

Suggested Citation

Wang, Yiyun and Liu, Zhenping and Wang, Xuchu and Dai, Yibei and Li, Xiaoying and Gao, Shifan and Yu, Pan and Lin, Qinyan and Fan, Zihao and Ping, Ying and Wang, Danhua and Lin, Xuedan and Zheng, Zejie and Liu, Weiwei and Tao, Zhihua, A Microfluidic ExoNANO for Efficient Isolation and Chemiluminescence Quantitative Immunoassay of Exosomes (April 12, 2018). Available at SSRN: https://ssrn.com/abstract=3296650 or http://dx.doi.org/10.2139/ssrn.3296650

Yiyun Wang

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Zhenping Liu

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Xuchu Wang

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Yibei Dai

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Xiaoying Li

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Shifan Gao

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Pan Yu

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Qinyan Lin

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Zihao Fan

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Ying Ping

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Danhua Wang

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Xuedan Lin

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Zejie Zheng

Zhejiang University

38 Zheda Road
Hangzhou, Zhejiang 310058
China

Weiwei Liu

Zhejiang University - Department of Laboratory Medicine ( email )

Hangzhou, 310009
China

Zhihua Tao (Contact Author)

Zhejiang University - Department of Clinical Laboratory ( email )

Zhejiang, Hangzhou
China

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