The group 2 allergen, Der p2, has been reported to activate innate toll-like receptors (TLRs) on respiratory epithelial cells and thus aggravate respiratory diseases. In this study, a high sensitive nanobiosensor based on a 3D sensing element that has uniformly deposited gold nanoparticles for the detection of the dust mite antigen Der p2 is proposed. The barrier layer of an anodic aluminum oxide (AAO) film is used as the template in this highly sensitive nanobiosensor fabricated with a reducing agent and stabilizer-free method. Electrochemical deposition is utilized to synthesize uniformly distributed gold nanoparticles on the surface of the barrier layer. The size and the distribution density of the nanoparticles can be well controlled by the potential applied during electrochemical deposition. Following this procedure, monoclonal antibodies were immobilized against the dust mite antigen Der p2 by the gold nanoparticles through the 11-MUA (11-mercaptoundecanoic acid), EDC (1-Ethyl-3-(3-dimethyl-aminopropyl)-carbodiimide)/NHS (N-hydroxysuccinimide) self-assembled monolayer approach. The proposed nanobiosensor was successfully used to examine the Der p2 down to a concentration of 1pg/mL through the electrochemical impedance spectroscopy analysis. The high sensitivity of the proposed 3D nanobiosensor can be attributed to the high intensity and uniformity of the Au nanoparticles on the sensor. The proposed nanobiosensor would be useful for the fast detection of rare molecules in a solution.

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