MOE Key Laboratory of Laser Life Science, Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, 510631, China; Guangdong Provincial Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, 510631, China; Guangzhou Key Laboratory of Spectral Analysis and Functional Probes, College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
MOE Key Laboratory of Laser Life Science, Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, 510631, China; Guangdong Provincial Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, 510631, China; Guangzhou Key Laboratory of Spectral Analysis and Functional Probes, College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
Anal Chim Acta. 2021 Dec 15;1188:339172. doi: 10.1016/j.aca.2021.339172. Epub 2021 Oct 20.
Tyrosinase (TYR) is as a well-known polyphenol oxidase and important biomarker of melanocytic lesions. Thus, developing powerful methods to determine TYR activity is of great value in the early diagnosis of skin disease. Direct surface-enhanced Raman scattering (SERS) detection of biomolecules is usually affected by non-specific interference and complicate structure of the analytes. It is a challenge to develop Raman-active molecules with specific recognition to analytes in complex media. Here, we report a novel colorimetric and surface-enhanced Raman scattering (SERS) dual-readout assay for the determination of TYR using commercially available and economical 4-mercaptophenyl boronic acid (4-MPBA) as a Raman-active and recognition molecule. 4-MPBA provides a unique interactive boronic acid group to the diol group of TYR substrate and exhibits good SERS signal. Also, the introduction of magnetic beads could promptly improve the anti-interference ability of dual-mode sensor. The TYR-incubated tyramine-modified magnetic beads could obviously change the concentration of 4-MPBA-AuNPs in the presence of O and ascorbic acid, where the ultraviolet visible (UV-vis) absorption and SERS intensity were directly related to the concentration of TYR added. The dual-mode sensor had a rapid response to TYR within 1 min under optimized conditions and had high selectivity for TYR with a limit of detection at 0.001 U/mL. In addition, the dual-mode strategy showed promising prospects in the determination of TYR activity in serum samples and could be used to screen TYR inhibitors.
酪氨酸酶（TYR）是一种众所周知的多酚氧化酶，也是黑素细胞病变的重要生物标志物。因此，开发强大的方法来测定 TYR 活性对于皮肤疾病的早期诊断具有重要价值。生物分子的直接表面增强拉曼散射（SERS）检测通常受到非特异性干扰和分析物复杂结构的影响。在复杂介质中开发具有对分析物具有特异性识别的拉曼活性分子是一个挑战。在这里，我们报告了一种使用市售且经济的 4-巯基苯硼酸（4-MPBA）作为拉曼活性和识别分子的新型比色和表面增强拉曼散射（SERS）双读取测定法，用于测定 TYR。4-MPBA 为 TYR 底物的二醇基团提供了独特的相互作用硼酸基团，并表现出良好的 SERS 信号。此外，引入磁性珠可以迅速提高双模传感器的抗干扰能力。TYR 孵育的酪胺修饰的磁性珠在 O 和抗坏血酸存在的情况下，可明显改变 4-MPBA-AuNPs 的浓度，其中紫外可见（UV-vis）吸收和 SERS 强度与添加的 TYR 浓度直接相关。在优化条件下，双模态传感器对 TYR 的快速响应时间为 1 分钟，对 TYR 具有高选择性，检测限为 0.001 U/mL。此外，双模态策略在血清样品中 TYR 活性的测定中显示出有前景的应用，可用于筛选 TYR 抑制剂。
