Fe3O4/Au magnetic nanoparticle amplification strategies for ultrasensitive electrochemical immunoassay of alfa-fetoprotein

Abstract

Ning Gan1*, Haijuan Jin1*, Tianhua Li1, Lei Zheng21The State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 2Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, People's Republic of China *Both authors contributed equally to this workBackground: The purpose of this study was to devise a novel electrochemical immunosensor for ultrasensitive detection of alfa-fetoprotein based on Fe3O4/Au nanoparticles as a carrier using a multienzyme amplification strategy.Methods and results: Greatly enhanced sensitivity was achieved using bioconjugates containing horseradish peroxidase (HRP) and a secondary antibody (Ab2) linked to Fe3O4/Au nanoparticles (Fe3O4/Au-HRP-Ab2) at a high HRP/Ab2 ratio. After a sandwich immunoreaction, the Fe3O4/Au-HRP-Ab2 captured on the electrode surface produced an amplified electrocatalytic response by reduction of enzymatically oxidized hydroquinone in the presence of hydrogen peroxide. The high content of HRP in the Fe3O4/Au-HRP-Ab2 could greatly amplify the electrochemical signal. Under optimal conditions, the reduction current increased with increasing alfa-fetoprotein concentration in the sample, and exhibited a dynamic range of 0.005–10 ng/mL with a detection limit of 3 pg/mL.Conclusion: The amplified immunoassay developed in this work shows good precision, acceptable stability, and reproducibility, and can be used for detection of alfa-fetoprotein in real samples, so provides a potential alternative tool for detection of protein in the laboratory. Furthermore, this immunosensor could be regenerated by simply using an external magnetic field.Keywords: Fe3O4/Au nanoparticles, alfa-fetoprotein, sandwich immunoassay, electrochemical immunosensor