Optimization of Nanowell-Based Label-Free Impedance Biosensor Based on Different Nanowell Structures

Abstract

Nanowell-based impedance-based label-free biosensors have demonstrated significant advantages in sensitivity, simplicity, and accuracy for detecting cancer biomarkers and macromolecules compared to conventional impedance-based biosensors. Although nanowell arrays have previously been employed for biomarker detection, a notable limitation exists in the photolithography step of their fabrication process, leading to a reduced efficiency rate. Historically, the diameter of these nanowells has been 2 μm. To address this issue, we propose alternative geometries for nanowells that feature larger surface areas while maintaining a similar circumference, thereby enhancing the fabrication efficiency of the biosensors. We investigated three geometries: tube, spiral, and quatrefoil. Impedance measurements of the samples were conducted at 10 min intervals using a lock-in amplifier. The study utilized interleukin-6 (IL-6) antibodies and antigens/proteins at a concentration of 100 nM as the target macromolecules. The results indicated that tube-shaped nanowells exhibited the highest sensitivity for detecting IL-6 protein, with an impedance change of 9.55%. In contrast, the spiral, quatrefoil, and circle geometries showed impedance changes of 0.91%, 0.95%, and 1.62%, respectively. Therefore, the tube-shaped nanowell structure presents a promising alternative to conventional nanowell arrays for future studies, potentially enhancing the efficiency and sensitivity of biosensor fabrication.