Aptamer-based fluorescence polarization assay for separation-free exosome quantification.

Zhen Zhang, Chuanhao Tang, Libo Zhao, Li Xu, Wei Zhou, Zaizai Dong, Yuqing Yang, Qiqi Xie, Xiaohong Fang
Author Information
  1. Zhen Zhang: CAS Key Laboratory of Molecular Nanostructure and Nanotechnology, Beijing National Research Center for Molecular Sciences, CAS Research/Education Center for Excellence in Molecule Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. xfang@iccas.ac.cn zhangzhen@iccas.ac.cn. ORCID
  2. Chuanhao Tang: Department of Medical Oncology, Peking University International Hospital, Beijing 102206, P. R. China.
  3. Libo Zhao: Echo Biotech Co. Ltd, Beijing 102206, P. R. China.
  4. Li Xu: CAS Key Laboratory of Molecular Nanostructure and Nanotechnology, Beijing National Research Center for Molecular Sciences, CAS Research/Education Center for Excellence in Molecule Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. xfang@iccas.ac.cn zhangzhen@iccas.ac.cn.
  5. Wei Zhou: CAS Key Laboratory of Molecular Nanostructure and Nanotechnology, Beijing National Research Center for Molecular Sciences, CAS Research/Education Center for Excellence in Molecule Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. xfang@iccas.ac.cn zhangzhen@iccas.ac.cn and University of Chinese Academy of Sciences, Beijing 100049, P. R. China.
  6. Zaizai Dong: CAS Key Laboratory of Molecular Nanostructure and Nanotechnology, Beijing National Research Center for Molecular Sciences, CAS Research/Education Center for Excellence in Molecule Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. xfang@iccas.ac.cn zhangzhen@iccas.ac.cn and University of Chinese Academy of Sciences, Beijing 100049, P. R. China.
  7. Yuqing Yang: Echo Biotech Co. Ltd, Beijing 102206, P. R. China.
  8. Qiqi Xie: Echo Biotech Co. Ltd, Beijing 102206, P. R. China.
  9. Xiaohong Fang: CAS Key Laboratory of Molecular Nanostructure and Nanotechnology, Beijing National Research Center for Molecular Sciences, CAS Research/Education Center for Excellence in Molecule Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. xfang@iccas.ac.cn zhangzhen@iccas.ac.cn and University of Chinese Academy of Sciences, Beijing 100049, P. R. China. ORCID

Abstract

Tumor-derived exosomes have emerged as promising cancer biomarkers and attracted increasing interest in non-invasive cancer diagnosis and treatment monitoring. However, the identification and quantification of exosomes in clinical samples such as blood remains challenging due to the difficulty in trade-off between recognition specificity and isolation efficiency. Here we have developed an aptamer-based fluorescence polarization assay for exosome quantification, which is a separation-free, amplification-free and sensitive approach enabling direct quantification of exosomes in human plasma. While the key specificity of this assay is based on the aptamer's inherent affinity to membrane proteins on exosomes, exosomes' inherent huge mass/volume acts as mass-based fluorescence polarization amplifier. Our assay allows quantitative analysis of exosomes in the range of 5 × 10 to 5 × 10 particles per μL with a detect limitation of 500 particles per μL for the cell line deprived exosomes. The total assay time is about 30 min with just one mix-and-read step to achieve high sensitivity. We have also demonstrated quantification of exosomes from lung cancer patients and healthy donors in clinical samples. This work describes a new and simple liquid biopsy assay to directly detect exosomes in the biological matrix, which facilitates cancer diagnosis and therapy monitoring.

MeSH Term

A549 Cells
Aptamers, Nucleotide
Biosensing Techniques
Exosomes
Fluorescence Polarization
Humans
Lung Neoplasms
Tetraspanin 30

Chemicals

Aptamers, Nucleotide
Tetraspanin 30

Word Cloud

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