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Triple-color fluorescence co-localization of PD-L1-overexpressing cancer exosomes

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Abstract

Programed cell death ligand 1 (PD-L1) is a protein biomarker overexpressed on exosomes derived from tumor cells. It plays an important role in tumor diagnosis, screening, evaluation of therapeutic efficacy, and prognosis. In this study, a facile method is presented to detect PD-L1-overexpressing cancer exosomes with high specificity and sensitivity. First, gold nanospheres (GNSs) were attached to the bottom of an eight-well chambered slide by electrostatic adsorption, forming the detection substrate. Then, Cy5-labeled CD63 aptamers (i.e., the capture probes) were modified on the GNSs by Au–S bond. After adding samples containing target exosomes which were stained by membrane dyes DiI in advance, FAM-labeled PD-L1 aptamers (i.e., the immunoprobes) were added to recognize PD-L1 on the target exosomes. By triple-color fluorescence co-localization (TFC) of the Cy5, DiI, and FAM channels, highly sensitive and reliable detection of the PD-L1-overexpressing exosomes was achieved in the concentration range 7.78 × 101 to 7.78 × 104 particles/mL with a detection limit down to 6 particles/mL. The advantages of the proposed detection method include the following; first, the detection substrate is easy to prepare and convenient to clean. Second, the TFC strategy can completely exclude nonspecific reaction sites and thus significantly improves the accuracy. Such a facile and reliable detection method holds a great potential in exosome-based cancer theranostics.

Graphical abstract

In this paper, we proposed a triple-color fluorescence co-localization (TFC) strategy to significantly improve the reliability of exosome detection and the detection substrate is easy to prepare and convenient to clean. In addition, the LOD is down to 6 particles/mL, which is quite low compared with other detection methods

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Funding

This work was supported by the Natural Science Foundation of China (NSFC) (62175027, 62175030), the Natural Science Foundation of Jiangsu Province (BK20201261), the Zhishan Scholars Program of Southeast University (2242021R41151) and the National Key Basic Research Program of China (2015CB352002).

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  1. Advanced Photonics Center, Southeast University, Nanjing, 210096, Jiangsu, China

    Jinxiu Wei, Kai Zhu, Zengwei Chen, Zhaoyan Yang, Kuo Yang, Zhuyuan Wang, Shenfei Zong & Yiping Cui

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  1. Jinxiu Wei
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Wei, J., Zhu, K., Chen, Z. et al. Triple-color fluorescence co-localization of PD-L1-overexpressing cancer exosomes. Microchim Acta 189, 182 (2022). https://doi.org/10.1007/s00604-022-05278-6

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