Aptamer-Engineered Cu₂O Nanocubes as a Surface-Modulated Catalytic Optical Sensor for Lung Cancer Cell Detection

Herein, fine and homogeneous Cu₂O nanocubes are synthesized and sensitized with a hairpin-structured AS1411 aptamer for the establishment of a biosensor for lung cancer cell detection. The Apt-Cu₂O nanocubes feature a recognition function in identifying a cancer-associated surface nucleolin protein. The intrinsic reduction catalytic ability is also confirmed by the use of two benchmark substrates, methylene blue (MB) and 4-nitrophenol (4-NP). The aptamer grafting on Apt-Cu₂O nanocubes is able to greatly prevent nonspecific-protein binding and to show specificity toward the nucleolin protein. The specific binding resulting from nucleolin protein leads to less exposure of the active area of the Apt-Cu₂O nanocubes, so the catalytic ability of Apt-Cu₂O nanocubes is thus diminished. The modulated catalytic ability led to less generation of the reduced 4-AP product, and the change in absorption of 4-AP allows the quantification of the nucleolin protein with a detection limit of 0.47 nM. The as-developed biosensor is applied to the detection of nucleolin-overexpressed A549 lung cancer cells, presenting a sensitive detection limit down to 20 cells. This may be ascribed to the clustering of surface nucleolin protein in a lipid raft membrane of cancer cells, as evidenced by a notable binding of Apt-Cu₂O nanocubes on the cancer cell surface. Real human serum samples spiked with cancer cells were also investigated, and a recovery rate of 87 ± 2.4% for 20 extracted cells validates the surface-modulated Apt-Cu₂O nanocubes-based catalytic optical biosensor as a promising tool for the detection of circulating tumor cells. The establishment of the Apt-Cu₂O nanocubes may allow for further studies on their use as a potential theranostics tool for cancer therapy.