Ultrasensitive enzyme-free electrochemical immunosensor based on hybridization chain reaction triggered double strand...

An ultrasensitive enzyme-free electrochemical immunoassay was developed for detection of the fg/mL level carcinoembryonic antigen (CEA) by using a double strand DNA@Au nanoparticle (dsDNA@AuNP) tag and hexaammineruthenium(III) chloride (RuHex) as the electroactive indicator. The dsDNA@AuNP was synthesized by one-pot hybrid polymerization of dsDNA on initiator DNA modified AuNPs via hybridization chain reaction. The immunosensor was prepared by covalently cross-linking capture antibody on chitosan/AuNP nanocomposite modified glass carbon electrode. The AuNPs accelerated the electron transfer and led to high detection sensitivity. With a sandwich-type immunoreaction and a biotin-streptavidin affinity reaction, the dsDNA@AuNP tag was conjugated on the immunocomplex to bring a high amount of RuHex to the electrode surface via electrostatic interaction, resulting in an amplified electrochemical signal. Under optimal conditions, the proposed sensing platform showed a wide linear detection range from 10 fg/mL to 10 ng/mL along with a detection limit of 3.2 fg/mL for CEA. The immunosensor exhibited high sensitivity and good stability, showing a promising application in early cancer diagnosis and could be extended to sensitive electrochemical biosensing of other analytes.