College of Chemistry and Materials Science, Fujian Provincial Key Laboratory of Advanced Oriented Chemical Engineer, Fujian Key Laboratory of Polymer Materials, Engineering Research Center of Industrial Biocatalysis, Fujian Province Higher Education Institutes, Fujian Normal University, Fuzhou, Fujian, 350007, China.
College of Chemistry and Materials Science, Fujian Provincial Key Laboratory of Advanced Oriented Chemical Engineer, Fujian Key Laboratory of Polymer Materials, Engineering Research Center of Industrial Biocatalysis, Fujian Province Higher Education Institutes, Fujian Normal University, Fuzhou, Fujian, 350007, China.
Anal Chim Acta. 2022 Sep 22;1227:340302. doi: 10.1016/j.aca.2022.340302. Epub 2022 Aug 24.
Hyaluronidase expression is known to be upregulated in various pathological conditions. A method based on a combination of ratiometric surface-enhanced Raman scattering (SERS) and magnetic separation is described for the determination of hyaluronidase (HAase) activity. Gold nanospheres (AuNPs) functionalized by 4-mercaptophenylboronic acid (4-MPBA) form stable cyclic esters with diol on hyaluronic acid (HA) by the boronic acid group, while FeO@DTNB@Au modified with mercaptoethylamine (MEA) was used as a capture substrate to bind to the carboxyl group on the surface of HA, forming the "Au@4-MPBA@HA/FeO@DTNB@Au@MEA" "core-satellite" structure. When HAase is present, HA is enzymatically disrupted, resulting in the destruction of the "core-satellite" structure, the SERS signal of 4-MPBA is subsequently weakened. The gold shell in the substrate protects the 5,5'-Dithio bis-(2-nitrobenzoic acid) (DTNB) from the external environment, which makes it become an ideal internal standard (IS) molecule for subsequent calibration. Under optimal conditions, the I/I varied in the range of 10 - 10 U‧mL HAase activity, with a limit of detection (LOD) of 0.32 mU‧mL，below the level of HAase in normal human body fluids. This method has been successfully applied to the determination of HAase activity in urine and is expected to provide a new method in disease detection, especially in the non-invasive detection of bladder cancer.
透明质酸酶的表达已知在各种病理条件下上调。本文描述了一种基于比率表面增强拉曼散射（SERS）和磁分离相结合的方法，用于测定透明质酸酶（HAase）活性。经 4-巯基苯硼酸（4-MPBA）功能化的金纳米球（AuNPs）通过硼酸基团与透明质酸（HA）上的二醇形成稳定的环酯，而用巯基乙胺（MEA）修饰的 FeO@DTNB@Au 则用作捕获底物，与 HA 表面的羧基结合，形成“Au@4-MPBA@HA/FeO@DTNB@Au@MEA”“核-卫星”结构。当存在 HAase 时，HA 被酶促破坏，导致“核-卫星”结构被破坏，随后 4-MPBA 的 SERS 信号减弱。基底中的金壳保护 5,5'-二硫代双（2-硝基苯甲酸）（DTNB）免受外部环境的影响，使其成为后续校准的理想内标（IS）分子。在最佳条件下，I/I 在 10 - 10 U‧mL 的范围内变化，HAase 活性的检出限（LOD）为 0.32 mU‧mL，低于正常人体液中的 HAase 水平。该方法已成功应用于尿液中 HAase 活性的测定，有望为疾病检测提供一种新方法，特别是在膀胱癌的非侵入性检测中。
