C-reactive protein (CRP) level in blood is associated with the risk of developing cardiovascular events in higher-risk populations. We present a sandwich ELISA-like assay for the determination of CRP in blood by citicoline-bovine serum albumin (citicoline-BSA) conjugate and aptamer-functionalized gold nanoparticles (aptamer-AuNPs) nanozyme. The CRP in the blood sample was selectively adsorbed to the ELISA plate coated by citicoline-BSA, and then incubated with added aptamer-AuNPs. AuNPs exhibited peroxidase activity and oxidized 3,3'5,5'-tetramethylbenzidine from colorless to blue, achieving the measurement at 652 nm. The amplified signal increased linearly in a wide range from 0.1 to 200 ng mL-1 and with a detection limit of 8 pg mL-1. Finally, the method was further tested using rat blood from an isoproterenol-induced myocardial infarction experimental model to confirm its applicability. The developed method could directly determine CRP in blood sample after dilution with high accuracy and sensitivity. This method has many advantages, such as easiness to prepare materials, good stability between batches, high specificity, low detection limit, low-cost, easiness to operate with simple instruments, the most remarkable of which is its excellent lot-to-lot stability over the classical ELISA.
Keywords: Aptamer-functionalized gold nanoparticles; C-reactive protein; Citicoline-bovine serum albumin conjugate; Nanozyme; Sandwich ELISA-Like assay.
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