A Label-free "Lock-key" Fluorescence Aptasensing Based on Triplex-helix DNA and G-quadruplex for CA15-3 Detection

Herein, we designed a label-free fluorescent aptasensor based on triple-helix DNA and G-quadruplex for carbohydrate antigen (CA15-3) detection. The triplex-helix structure can be formed with inserted G-rich DNA (IG) and aptamer DNA (Apt), which like a "lock" to lock the G-rich sequences. The CA15-3 was the "key", which specifically combined with aptamer sequences of Apt, resulting in liberating IG from the triplex-helix "lock". Then, the G-rich sequences of IG were formed into G-quadruplex and specifically interacted with N-methylmesoporphyrin IX (NMM), which greatly enhanced the fluorescence of the solution. However, when the "key" did not exist, the "lock" was fastened and fluorescence intensity did not change. With this proposed method, the concentration of CA15-3 can be effectively detected from 0.01 to 5 U mL^-1 with a detection limit (LOD) of 0.01 U mL^-1. Furthermore, this proposed biosensor can be applied to spiked human serum with great precision and reproducibility.