Upconversion nanoparticle-based fluorescence resonance energy transfer sensing platform for the detection of cathepsin B activity in vitro and in vivo.

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: 980nm; emission: 541nm/655nm) and Cy5 (excitation: 645nm) 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 100ng·mL-1 in buffer and 2 × 103 to 1 × 105 cells in 0.2mL cell lysates. The limits of detection (LODs) are 0.30ng·mL-1 in buffer and 887 cells in 0.2mL of cell lysates (S/N = 3). The viability of USP to detect CTSB activity in tumor-bearing mice is hasfurtherbeen investigated using in vivo fluorescent imaging.