A ratiometric electrochemical sensor for the determination of exosomal glycoproteins

Abnormal glycosylation of exosomal proteins is related to many diseases. However, there is still a lack of convenient and easy methods for the determination of exosomal glycoproteins. In this work, a ratiometric electrochemical sensor based on the recognition of glycoproteins by boronic acid and core-shell nanoparticles of silica-silver (SiO₂@Ag) amplified signals was developed for the highly sensitive detection of exosomal glycoproteins. The CD63 aptamer-SiO₂-N-(2-((2-aminoethyl)disulfanyl)ethyl) ferrocene carboxamide (FcNHSSNH₂) probe was first connected to graphene oxide-cucurbit [7] (GO-CB [7]) modified GCE through host-guest recognition. The CD63 aptamer was employed for the specific capture of exosomes, and the FcNHSSNH₂ molecule was used as the internal reference signal of the sensor. The mercaptophenylboronic acid (MPBA) of MPBA-SiO₂@Ag probe was used for the identification of exosomes surface glycoproteins. SiO₂ nanoparticle has a large specific surface area, which can load a large amount of silver nanoparticles (AgNPs) for electrochemical signal amplification. The results were expressed as the current ratio of AgNPs and FcNHSSNH₂. The introduction of the internal reference molecule FcNHSSNH₂ could effectively reduce the measurement error caused by the different DNA density of the substrate, and further improve the sensitivity and accuracy of the detection. Under the optimal experimental conditions, this sensor allowed the sensitive detection of exosomal glycoproteins in the range of 4.2 × 10² to 4.2 × 10⁸ particles/μL with a limit of detection (LOD) of 368 particles/μL. Furthermore, the ratiometric electrochemical sensor could be employed for the detection of exosomal glycoproteins in human serum samples, which has a good clinical application prospect.