期刊
BIOSENSORS & BIOELECTRONICS
卷 26, 期 6, 页码 2905-2910出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2010.11.035
关键词
Aptasensor; Electrochemistry; Protein array; Intramolecular displacement
类别
资金
- National Nature Science Funding of China [90717104, 21075068]
- National Basic Research Program of China (973 Program) [2007CB914100, 2011CB707703]
- SKLEAC, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences [SKLEAC2010004]
Tripropylamine (TPA) has different oxidation efficiency at double stranded (ds)-and single stranded (ss)DNA-modified electrodes. Using this property, a simple but sensitive biosensor using TPA oxidation to probe the intramolecular displacement was constructed with the analysis of lysozyme as model for the first time. After the complementary ss-DNA strand of anti-lysozyme aptamer was immobilized onto gold electrode via gold-thiol bond, the incubation with the aptamer resulted in the formation of ds-DNA. Lysozyme (in 10 mu L sample) binding with aptamer displaced the complementary strand because of the high affinity of lysozyme and its aptamer, corresponding to the dissociation of the ds-DNA. The modified electrode was swept in 20 mM TPA solution from 0.2 to 0.95 V. The difference in oxidation current was used to quantify the content of lysozyme with a linear range from 1.0 pM to 1.1 nM. That means 10 amol or 6.0 x 10(6) lysozyme molecules can be detected. Because the signal is produced from the preconcentrated TPA at the electrode surface, the high sensitivity is achieved over the single site labelling strategy. The proposed method is simple, stable, specific, and time-saving while the complicated sample pre-treatment and the labelling to the DNA strand are avoided. The biosensor was validated by the analysis of the diluted egg white sample directly. The recovery and reproducibility were 93.3-100% and 1.4-4.2%, respectively. (C) 2010 Elsevier B.V. All rights reserved.
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