Enzyme-Free Electrochemical Immunosensor Based on Host–Guest Nanonets Catalyzing Amplification for Procalcitonin Detection

An enzyme-free electrochemical immunosensor based on the host–guest nanonets of N,N-bis(ferrocenoyl)-diaminoethane/β-cyclodextrins/poly(amidoamine) dendrimer-encapsulated Au nanoparticles (Fc-Fc/β-CD/PAMAM-Au) for procalcitonin (PCT) detection has been developed in this study. The signal probe was constructed as follows: amine-terminated β-CD was adsorbed to PAMAM-Au first, and then the prepared Fc-Fc was recognized by the β-CD to form stable host–guest nanonets. Next, secondary antibodies (Ab2) were attached into the formed netlike nanostructure of Fc-Fc/β-CD/PAMAM–Au by chemical absorption between PAMAM-Au and -NH2 of β-CD. Herein, the PAMAM-Au act not only as nanocarriers for anchoring large amounts of the β-CD and Ab2 but also as nanocatalysts to catalyze the oxidation of ascorbic acid (AA) for signal amplification. Moreover, the Fc-Fc could be stably immobilized by the hydrophobic inner cavity of β-CD as well as improving solubility by the hydrophilic exterior of β-CD. With the unique structure of two ferrocene units, Fc-Fc not only affords more electroactive groups to make the electrochemical response more sensitive but also plays a role of combining dispersive β-CD-functionalized PAMAM-Au to form the netlike nanostructure. Furthermore, Fc-Fc exhibits good catalytic activity for AA oxidation. When the detection solution contained AA, the synergetic catalysis of PAMAM-Au and Fc-Fc to AA oxidation could be obtained, realizing enzyme-free signal amplification. The proposed immunosensor provided a linear range from 1.80 pg/mL to 500 ng/mL for PCT detection and a detection limit of 0.36 pg/mL under optimal experimental conditions. Moreover, the immunosensor has shown potential application in clinical detection of PCT.