A smart TESTER for reliable discrimination of cancer-derived small extracellular vesicles

Ruyan Niu; Xiaohui Chen; Zixin Sun; Liu Wang; Zining Wang; Chong Zhang; Dan Ding; Jichun Yang; Yongzhong Wang; Yang Luo

doi:10.1016/j.aca.2023.341636

A smart TESTER for reliable discrimination of cancer-derived small extracellular vesicles

Anal Chim Acta. 2023 Oct 2:1276:341636. doi: 10.1016/j.aca.2023.341636. Epub 2023 Jul 17.

Authors

Ruyan Niu¹, Xiaohui Chen¹, Zixin Sun², Liu Wang², Zining Wang², Chong Zhang¹, Dan Ding³, Jichun Yang⁴, Yongzhong Wang⁵, Yang Luo⁶

Affiliations

¹ Key Laboratory for Biorheological Science and Technology of Ministry of Education, College of Bioengineering, NHC Key Laboratory of Birth Defects and Reproductive Health, Chongqing University, Chongqing, 400044, PR China; Center of Smart Laboratory and Molecular Medicine, School of Medicine, Chongqing University, Chongqing, 400044, PR China.
² Center of Smart Laboratory and Molecular Medicine, School of Medicine, Chongqing University, Chongqing, 400044, PR China.
³ College of Life Sciences, Nankai University, Tianjin, 300071, PR China.
⁴ Center of Smart Laboratory and Molecular Medicine, School of Medicine, Chongqing University, Chongqing, 400044, PR China. Electronic address: yangjichun@cqu.edu.cn.
⁵ Key Laboratory for Biorheological Science and Technology of Ministry of Education, College of Bioengineering, NHC Key Laboratory of Birth Defects and Reproductive Health, Chongqing University, Chongqing, 400044, PR China. Electronic address: wangyzh@cqu.edu.cn.
⁶ Key Laboratory for Biorheological Science and Technology of Ministry of Education, College of Bioengineering, NHC Key Laboratory of Birth Defects and Reproductive Health, Chongqing University, Chongqing, 400044, PR China; Center of Smart Laboratory and Molecular Medicine, School of Medicine, Chongqing University, Chongqing, 400044, PR China; College of Life Science and Laboratory Medicine, Kunming Medical University, Kunming, Yunnan, 650050, PR China. Electronic address: luoy@cqu.edu.cn.

PMID: 37573115
DOI: 10.1016/j.aca.2023.341636

Abstract

Cancer-derived small extracellular vesicles (csEVs) are crucial liquid biopsy indicators that reflect the presence and progression of many malignancies. However, reliable discrimination of csEVs remains a great challenge owing to the interference from normal sEVs (nsEVs) and low abundance in the early stages of cancer. In this work, we developed a Two-Elements Selectively Triggered csEVs Recognization (TESTER) strategy for selective identification of csEVs from the complex clinical body fluid samples. This method was based on the MNAzyme-controlled synchronous recognition to EpCAM and CD63 proteins on the membrane of csEVs. Efficient recognition to csEVs via EpCAM aptamer and CD63 aptamer prompted the release of Partzyme A and Partzyme B probes to induce a MNAzyme structure formation, resulting in the cyclic cleavage of substrate chain to produce cascade fluorescence signal amplification. The detection threshold of the developed TESTER approach for csEVs in complicated biological samples was 72 particles μL^-1, accomplishing the highly sensitive and selective quantification of csEVs. At the same time, we successfully constructed a new platform for bimolecular simultaneous recognition, which provides a good idea for the construction of bimolecular-activated detection switch in the future.

Keywords: Biosensor; Cancer; Detection; Extracellular vesicle; MNAzyme.