Nucleolin-Targeted Ratiometric Fluorescent Carbon Dots with a Remarkably Large Emission Wavelength Shift for Precise Imaging of Cathepsin B in Living Cancer Cells.

Abstract

As one of the most promising biomarkers for numerous malignant tumors, accurate and reliable reporting of Cathepsin B (CTSB) activity is of great significance to achieve efficient diagnosis of cancers at an early stage and predicting metastasis. Here, we report a vigorous ratiometric fluorescent method integrating a cancer-targeting recognition moiety with a remarkably large emission wavelength shift into a single matrix to report CTSB activity sensitively and specifically. As a proof of concept, we synthesized amine-rich carbon quantum dots (CQDs) with a blue fluorescence, which offered an efficient scaffolding to covalently assemble the nucleolin-targeting recognition nucleic acid aptamer AS1411 and a CTSB-cleavable peptide substrate Gly-Arg-Arg-Gly-Lys-Gly-Gly-Cys-COOH that tethered with a near-infrared (NIR) fluorophore chlorin e6 (Ce6-GRRGKGGC, Ce6-Pep), enabling a cancer-targeting and CTSB stimulus-responsive ratiometric nanoprobe AS1411-Ce6-CQDs. Owing to the efficient fluorescence resonance energy transfer (FRET) process from the CQDs to Ce6 inside the assembly of nanoprobe, the blue fluorescence of CQDs at ∼450 nm was remarkably quenched, along with an obvious NIR fluorescence enhancement of Ce6 at ∼650 nm. After selective entry into cancer cells via nucleolin-mediated endocytosis, the overexpressed CTSB in lysosome could cleave Ce6-Pep and trigger the Ce6 moiety dissociation from AS1411-Ce6-CQDs, thus leading to the termination of FRET process, achieving the efficient ratiometric fluorescence response toward endogenous CTSB with a remarkably large emission wavelength shift of ∼200 nm from NIR to blue emission region. Notably, the nanoprobe AS1411-Ce6-CQDs exhibited an excellent specificity for ratiometric fluorescent sensing of CTSB activity with an ultralow detection limit of 0.096 ng/mL, demonstrating its promising use for early precise cancer diagnosis in the near future.

• Authors: Yizhong Shen, Tingting Wu, Yuqi Wang, Shao-Lin Zhang, Xueli Zhao, Hong-Yuan Chen, Jing-Juan Xu
• Journal: Analytical chemistry (Anal Chem) Vol. 93 Issue 8 Pg. 4042-4050 (03 02 2021) ISSN: 1520-6882 [Electronic] United States
• PMID: 33586959 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Phosphoproteins
• RNA-Binding Proteins
• nucleolin
• Carbon
• Cathepsin B

Topics

• Carbon
• Cathepsin B
• Fluorescence Resonance Energy Transfer
• Neoplasms (diagnostic imaging)
• Phosphoproteins
• Quantum Dots
• RNA-Binding Proteins