Tyrosinase (TYR) is a polyphenol oxidase that regulates melanin biosynthesis . Abnormal levels of TYR have been confirmed closely associated with melanoma cancer and other diseases, making the establishment of highly sensitive and accurate quantitative detection of TYR is thus essential for fundamental research and clinical applications. Herein, we proposed a new strategy that combines surface-enhanced Raman scattering (SERS) with Dopamine (DA) and Prussian blue (PB) functionalized gold-gold hybrid nanoparticles for TYR detection. DA is oxidized to dopaquinone with the presence of TYR, leading to the consumption of DA in the reaction solution and the corresponding decrease in DA characteristic peak intensity at 1480 cm −1 . Our SERS quantitative assay of TYR with “on-off” sensing strategy yields an excellent limit of detection (LOD) of 3 × 10 −3 U mL −1 in a linear range of 10 −3 to 100 U mL −1 . In particular, the intensity ratio of 1480 cm −1 to 2121 cm −1 allows us to achieve remarkably reliable quantitative detection of TYR, with the 2121 cm −1 peak intensity of the standard internal (IS) molecule PB being used to correct SERS signal fluctuations. Furthermore, our proposed assay has been successfully demonstrated to quantify TYR spiked in human serum samples, with excellent percentage recovery of 90.0–110.6 %. The potential of our ratiometric SERS strategy for the performance evaluation of TYR inhibitors has also been verified. Our work is therefore expected to provide a new route for accurate and reliable monitoring of TYR activity in the complex biological environment.
Minling Li,Dechan Lu,Ruiyun You,Huiying Shen,Lanjin Zhu,Qingqiang Lin,Yudong Lu
Tyrosinase (TYR) is a key rate-limiting enzyme of melanin and a biomarker for melanoma diagnosis. Therefore, detection of TYR in biological samples is essential for the clinical diagnosis of melanoma. A novel portable surface-enhanced Raman scattering (SERS) sensor is proposed to detect TYR activity. It is based on dopamine (DA)-functionalized Au nanorod (Au NR) arrays as the capture substrate and 4-mercaptophenylboronic acid (4-MPBA)-modified silver nanoparticles (Ag NPs) as the SERS probe. In this case, the Au NR arrays are modified with 3,3′-dithiodipropionic acid di(N-succinimide ester) (DSP) to capture DA. In the presence of TYR, DA is oxidized to dopaquinone (DQ), which resulted in a decrease in the concentration of SERS probes further adsorbed onto the arrays. The activity of TYR is detected by changes in the intensity of the phenylboronic acid signal. Furthermore, this detection system has a wide linear detection range and high sensitivity that can effortlessly evaluate a linear range of 0.0001–100 U/mL of TYR activity. More significantly, the presented SERS sensor can perform well on both the determination of TYR activity in serum and the assessment of TYR inhibitors, which offers new opportunities for diagnosis of melanoma and other TYR-related diseases under complex biological conditions.
危险品化学品经营许可证（带存储）