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Signal-On Dual-Potential Electrochemiluminescence Based on Luminol–Gold Bifunctional Nanoparticles for Telomerase Detection

Zhang, Huai-Rong, Wu, Mei-Sheng, Xu, Jing-Juan, Chen, Hong-Yuan
Analytical chemistry 2014 v.86 no.8 pp. 3834-3840
apoptosis, complementary DNA, electrochemiluminescence, glassy carbon electrode, hydrogen peroxide, nanocrystals, nanogold, nanoparticles, neoplasm cells, surface plasmon resonance, telomerase
Here, we report a novel type of signal-on dual-potential electrochemiluminescence (ECL) approach for telomerase detection based on bifunctionalized luminol–gold nanoparticles (L–Au NPs). In this approach, CdS nanocrystals (NCs) were first coated on glassy carbon electrode, and then thiol-modified telomerase primer was attached on CdS NCs via Cd–S bond. In the presence of telomerase and dNTPs, the primer could be extended. Telomerase primer would hybridize with its complementary DNA, and the extended part would hybridize with the capture DNA which was tagged with L–Au NPs. In the presence of coreactant H₂O₂, the L–Au NPs could not only enhance the ECL intensity of CdS NCs at −1.25 V (vs SCE) induced by the surface plasmon resonance (SPR) of Au NPs but also produce a new ECL signal at +0.45 V (vs SCE) that resulted from luminol in L–Au NPs. Both signals at two potentials increased with the increase of telomerase concentration. This method could be used to detect the telomerase from 100 to 9000 HL-60 cells and investigate the apoptosis of tumor cells. The ratio of the two signal increments (ΔECLLᵤₘᵢₙₒₗ/ΔECLCdS NCₛ), which showed a high consistency value for different numbers of cells, could be used to verify the reliability of tests. This dual-potential ECL strategy showed great promise in avoiding false positive or negative results in bioanalysis.