Abstract
A simple fluorescence resonance energy transfer (FRET) sensing platform (termed as USP), comprised of upconversion nanoparticles (UCNPs) as the energy donor and Cy5 as the energy acceptor, has been synthesized for cathepsin B (CTSB) activity detection in vitro and in vivo. When Cy5-modified peptide substrate (peptide-Cy5) of CTSB is covalently linked on the surface of UCNPs, the FRET between the UCNPs (excitation: 980 nm; emission: 541 nm/655 nm) and Cy5 (excitation: 645 nm) leads to a reduction in the red upconversion luminescence (UCL) signal intensity of UCNPs. Cy5 can be liberated from UCNPs in the presence of CTSB through the cleavage of peptide-Cy5 by CTSB, leading to the recovery of the red UCL signal of UCNPs. Because the green UCL signal of UCNPs remains constant during the CTSB digestion, it can be considered as an internal reference. The findings demonstrate the ability of USP to detect CTSB with the linear detection ranges of 1 to 100 ng·mL−1 in buffer and 2 × 103 to 1 × 105 cells in 0.2 mL cell lysates. The limits of detection (LODs) are 0.30 ng·mL−1 in buffer and 887 cells in 0.2 mL of cell lysates (S/N = 3). The viability of USP to detect CTSB activity in tumor-bearing mice is has further been investigated using in vivo fluorescent imaging.
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The data that support the findings of this study are available from the corresponding author upon reasonable request.
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Funding
The authors would like to thank the National Natural Science Foundation of China (Grant No. 81871406 and 61901438) and the Achievement Transformation Guidance Fund Between the First Hospital of Jilin University and Changchun Institute of Applied Chemistry (Grant No. CGZHYD202012-009) for financial support.
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Li, X., Zhang, M., Zhang, H. et al. Upconversion nanoparticle-based fluorescence resonance energy transfer sensing platform for the detection of cathepsin B activity in vitro and in vivo. Microchim Acta 190, 181 (2023). https://doi.org/10.1007/s00604-023-05771-6
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DOI: https://doi.org/10.1007/s00604-023-05771-6