Dually enhanced homogenous synthesis of molybdophosphate by hybridization chain reaction and enzyme nanotags for the electrochemical bioassay of carcinoembryonic antigen

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.