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Updated: Apr 24, 2026
Fabrication of Electrochemical-DNA Biosensors for the Reagentless Detection of Nucleic Acids, Proteins and Small Molecules
Published on: June 1, 2011
Fabricating a reversible and regenerable electrochemical biosensor for quantitative detection of antibody by using
Wei Wei1, Linqun Zhang1, Qiangwei Ni1
1Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Nanjing 211189, Jiangsu, PR China.
Abstract:
A reversible and regenerable electrochemical biosensor is fabricated for quantitative detection of antibody based on "triplex-stem" molecular switches. A hairpin-shaped oligonucleotide (hairpin DNA) labeled with ferrocene (Fc) at the 3'-end is fixed on the gold electrode serving as a signal transduction probe. Its hairpin structure leads Fc close to the surface of gold electrode and produces a strong current signal (on-state). A single-strand oligonucleotide modified with two digoxin molecules on the two arm segments (capture DNA) interact with hairpin DNA with the help of Ag(+) ions. The "triplex-stem" DNA forms, which separates Fc from the electrode and reduces the electrochemical signal (off-state). Binding of digoxin antibody to digoxin releases capture DNA from the hairpin DNA, creating an effective "off-on" current signal switch. The stability of the "triplex-stem" structure of hairpin/capture DNA is critical to the signal switch and the sensitivity of the method, which can be adjusted conveniently and efficiently by changing Ag(+) concentrations. Based on the "off-on" current signal switch, this biosensor is used to detect digoxin antibody sensitively in blood serum. The linear range is 1.0-500 pg with a correlation coefficient of 0.996, and the detection limit is 0.4 pg. Also, this biosensor shows excellent reversibility and reproducibility, which are significant requirements for practical biosensor applications.
