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Int. J. Mol. Sci. 2011, 12(11), 7410-7423; doi:10.3390/ijms12117410

A Sandwich Electrochemical Immunosensor Using Magnetic DNA Nanoprobes for Carcinoembryonic Antigen

1,†,* , 1,2,†
and
2,*
1
The State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, Zhejiang, China
2
Clinical Laboratory Center, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong, China
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Received: 11 July 2011 / Revised: 23 September 2011 / Accepted: 21 October 2011 / Published: 28 October 2011
(This article belongs to the Section Material Sciences and Nanotechnology)
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Abstract

A novel magnetic nanoparticle-based electrochemical immunoassay of carcinoembryonic antigen (CEA) was designed as a model using CEA antibody-functionalized magnetic beads [DNA/Fe3O4/ZrO2; Fe3O4 (core)/ZrO2 (shell) nano particles (ZMPs)] as immunosensing probes. To design the immunoassay, the CEA antibody and O-phenylenediamine (OPD) were initially immobilized on a chitosan/nano gold composite membrane on a glassy carbon electrode (GCE/CS-nano Au), which was used for CEA recognition. Then, horseradish peroxidase (HRP)-labeled anti-CEA antibodies (HRP-CEA Ab2) were bound to the surface of the synthesized magnetic ZMP nanoparticles as signal tag. Thus, the sandwich-type immune complex could be formed between secondary antibody (Ab2) modified DNA/ZMPs nanochains tagged by HRP and GCE/CS-nano Au. Unlike conventional nanoparticle-based electrochemical immunoassays, the recognition elements of this immunoassay included both electron mediators and enzyme labels, which obviously simplifies the electrochemical measurement process. The sandwich-type immunoassay format was used for online formation of the immunocomplex of CEA captured in the detection cell with an external magnet. The electrochemical signals derived from HRP during the reduction of H2O2 with OPD as electron mediator were measured. The method displayed a high sensitivity for CEA detection in the range of 0.008–200 ng/mL, with a detection limit of 5 pg/mL (estimated at a signal-to-noise ratio of 3). The precision, reproducibility, and stability of the immunoassay were good. The use of the assay was evaluated with clinical serum samples, and the results were in excellent accordance with those obtained using the standard enzyme-linked immunosorbent assay (ELISA) method. Thus, the magnetic nanoparticle-based assay format is a promising approach for clinical applications, and it could be further developed for the detection of other biomarkers in cancer diagnosis.
Keywords: chitosan/DNA/Fe3O4/ZrO2; magnetic nanoparticle-based electrochemical immunoassay; carcinoembryonic antigen; sandwich electrochemical immunosensor chitosan/DNA/Fe3O4/ZrO2; magnetic nanoparticle-based electrochemical immunoassay; carcinoembryonic antigen; sandwich electrochemical immunosensor
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Gan, N.; Jia, L.; Zheng, L. A Sandwich Electrochemical Immunosensor Using Magnetic DNA Nanoprobes for Carcinoembryonic Antigen. Int. J. Mol. Sci. 2011, 12, 7410-7423.

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