Hybridization chain reaction-enhanced enzyme biomineralization for ultrasensitive colorimetric biosensing of a protein biomarker

Abstract

By employment of an aptamer-initiated hybridization chain reaction (HCR) to enhance the enzyme biomineralization of cupric subcarbonate, this work develops a novel colorimetric biosensing method for protein analysis. The HCR product was used to specifically attach a large amount of urease-functionalized gold nanoparticles (Au NPs) for the preparation of a gold nanoprobe. After the sandwich biorecognition reactions, this nanoprobe could be quantitatively captured onto the antibody-functionalized magnetic bead (MB) platform. Then, numerous copper ions would be enriched onto the MB surface through the urease-induced biomineralization of cupric subcarbonate. Based on the complete release of Cu²⁺ ions for the sensitive copper chromogenic reaction, convenient colorimetric signal transduction was thus achieved for the quantitative analysis of the target analyte of the carcinoembryonic antigen. The HCR product provides a large number of biotin sites for the attachment of Au NP nanotags. The biomineralization reaction of high-content urease loaded onto Au NPs leads to highly efficient Cu²⁺ enrichment for signal amplification. So this method features excellent performance including a very wide linear range and a low detection limit down to 0.071 pg mL⁻¹. In addition, the satisfactory results of real sample experiments reveal that this method possesses huge potential for practical applications.