Label-Free and Regenerable Aptasensor for Real-Time Detection of Cadmium(II) by Dual Polarization Interferometry

Recently, numerous aptamer-based biosensors have been developed to detect heavy metal ions. Most of aptamer-based biosensors only can be used to get some quantitative information. The mechanism of the interaction between aptamer and metal ions, however, is rarely studied. In this work, a label-free and regenerable aptamer-based biosensor was constructed using dual polarization interferometry (DPI). This aptasensor was used to investigate the real-time interaction process between cadmium(II) and its aptamer. According to the information on mass, thickness, and density obtained by DPI, a Cd²⁺ concentration-dependent interaction mechanism and conformation of aptamer was proposed. At low Cd²⁺ concentration, Cd²⁺ mainly interacted with phosphate groups on aptamer, resulting in the stretched ssDNA and a few vertical hairpin structures. When adding the high concentration of Cd²⁺, Cd²⁺ primarily bound with bases of DNA by coordination interaction and the conformation of aptamer transferred to a tight and short hairpin structure. In addition, the association rate constant (k_a), dissociation rate constant (k_d) and disassociation constant (K_D) between Cd²⁺ and its aptamer were calculated to be 96 M^-1 S^-1, 2.11 × 10^-5 S^-1, and 220 nM, respectively. The proposed aptasensor showed high sensitivity for Cd²⁺ detection with the detection limit of 0.61 μg/L, which was far below the 5.0 μg/L ranked by the U.S. Environmental Protection Agency. The biosensor also exhibited excellent regenerability and could be used for three cycles without obvious change in response signal. Therefore, the developed method could not only provide quantitative information, but also offered the information on conformation and kinetics for molecular interaction. This method can facilely extend to study the interaction of DNA with other ions, small molecules, or biomacromolecules.