Citation: ZHU Hua-Yu, ZHANG Li, CHEN Huai-Cheng, YAN Sheng-Juan. Electrochemiluminescence Biosensor for Detection of Thrombin Using Enzyme-based Signal Amplification[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(10): 1549-1554. doi: 10.3724/SP.J.1096.2012.11394
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Amino functionalized gold nanoparticles(GNP) were synthesized in this work. Electrochemiluminescence (ECL) probe was synthesized by covalently linking tris (2,2-bipyridyl) ruthenium (Ⅱ) and self-assembly of thiol DNA5 to this GNP. By using nicking endonuclease signal amplification technology, large amount of newly obtained DNA fragments was obtained to capture ECL probe. Thiol DNA probe was self-assembled on gold electrode surface, and then ECL biosensor was fabricated by sequentially hybridization of complementary DNA (newly obtained DNA fragment) and ECL probe to this modified gold electrode. Under optimized conditions, the ECL intensity of this biosensor exhibited linear relationship with thrombin concentration in the range of 3.0×10-13-6.0×10-11 mol/L, and the detection limit was 1.8×10-13 mol/L (3σ). The biosensor shows good application prospect in the field of biological samples analysis owing to its high sensitivity, good stability and reproducibility.
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