A magnetic bead (MB)-based sandwich biorecognition reactions is combined with a gold nanoprobe-induced homogenous synthesis of molybdophosphate to develop a novel bioassay method for the electrochemical detection of the tumor biomarker of carcinoembryonic antigen (CEA). The nanoprobe is prepared through the specific loading of numerous alkaline phosphatase (ALP)-functionalized gold nanoparticles (Au NPs) on a double-stranded DNA (dsDNA) produced by the CEA aptamer-triggered hybridization chain reaction (HCR). Both the large amounts of PO43− produced by the ALP catalytic hydrolysis of pyrophosphate and the phosphate backbones of dsDNA can react with the added MoO42− to generate electroactive molybdophosphates. So, the gold nanoprobe was used for signal tracing of the sandwich bioassay of CEA at a constructed antibody-functionalized MB platform. The sensitive electrochemical measurement of molybdophosphate produced from the quantitatively captured nanoprobes at a carbon nanotube-modified electrode (measured at about 0.12 V vs. Ag/AgCl, 3 M KCl) enabled the convenient signal transduction of the method. Due to the dually enhanced synthesis of molybdophosphate by the HCR and multi-enzyme Au NP nanotags, this method shows a wide linear range from 0.05 pg mL−1 to 10 ng mL−1 along with a low detection limit of 0.027 pg mL−1. In addition, the MB-based biorecognition reaction and the homogeneous synthesis of molybdophosphate are much convenient in manipulations. These excellent performances decide the extensive application potentials of the method.
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This work was financially supported by the National Natural Science Foundation of China (21475033) and Science and Technology Foundation for Excellent Creative Research Group of Hubei Provincial Department of Education (T201810).
All serum sample experiments were approved by the Ethics Committee of Hubei Normal University, and informed consent was obtained from human participants of this study.
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Cai, X., Lv, F., Lai, G. et al. Dually enhanced homogenous synthesis of molybdophosphate by hybridization chain reaction and enzyme nanotags for the electrochemical bioassay of carcinoembryonic antigen. Microchim Acta 187, 361 (2020). https://doi.org/10.1007/s00604-020-04342-3
- Protein biomarker
- Hybridization chain reaction
- Electroanalytical chemistry