Cancer Research Center& Jiangxi Engineering Research Center for Translational Cancer Technology, Jiangxi University of Chinese Medicine, Nanchang, 330004, Jiangxi, China.
Institute for Advanced Study, Research Center for Differentiation and Development of TCM Basic Theory, Jiangxi University of Chinese Medicine, Nanchang, 330004, Jiangxi, China.
Mikrochim Acta. 2024 Jan 17;191(2):101. doi: 10.1007/s00604-024-06181-y.
Hyaluronidase (HAase) is an important endoglycosidase involved in numerous physiological and pathological processes, such as apoptosis, senescence, and cancer progression. Simple, convenient, and sensitive detection of HAase is important for clinical diagnosis. Herein, an easy-to-operate multicolor visual sensing strategy was developed for HAase determination. The proposed sensor was composed of an enzyme-responsive hydrogel and a nanochromogenic system (gold nanobipyramids (AuNBPs)). The enzyme-responsive hydrogel, formed by polyethyleneimine-hyaluronic acid (PEI-HA), was specifically hydrolyzed with HAase, leading to the release of platinum nanoparticles (PtNPs). Subsequently, PtNPs catalyzed the mixed system of 3,3',5,5'-tetramethylbenzidine (TMB) and HO to produce TMB under acidic conditions. Then, TMB effectively etched the AuNBPs and resulted in morphological changes in the AuNBPs, accompanied by a blueshift in the localized surface plasmon resonance peak and vibrant colors. Therefore, HAase can be semiquantitatively determined by directly observing the color change of AuNBPs with the naked eye. On the basis of this, the method has a linear detection range of HAase concentrations between 0.6 and 40 U/mL, with a detection limit of 0.3 U/mL. In addition, our designed multicolor biosensor successfully detected the concentration of HAase in human serum samples. The results showed no obvious difference between this method and enzyme-linked immunosorbent assay, indicating the good accuracy and usability of the suggested method.
透明质酸酶(HAase)是一种参与多种生理和病理过程的重要内切糖苷酶,如细胞凋亡、衰老和癌症进展。简单、方便、灵敏地检测 HAase 对于临床诊断非常重要。本文开发了一种易于操作的多色可视化传感策略来测定 HAase。该传感器由酶响应水凝胶和纳米显色系统(金纳米双锥体(AuNBPs))组成。酶响应水凝胶由聚乙烯亚胺-透明质酸(PEI-HA)形成,可特异性地被 HAase 水解,导致铂纳米颗粒(PtNPs)的释放。随后,PtNPs 在酸性条件下催化 TMB 和 HO 的混合体系产生 TMB。然后,TMB 有效地刻蚀 AuNBPs,导致 AuNBPs 的形态发生变化,同时局域表面等离子体共振峰发生蓝移并呈现出鲜艳的颜色。因此,可以通过肉眼直接观察 AuNBPs 的颜色变化来对半定量地测定 HAase。在此基础上,该方法对 HAase 浓度在 0.6 至 40 U/mL 之间具有线性检测范围,检测限为 0.3 U/mL。此外,我们设计的多色生物传感器成功地检测了人血清样品中 HAase 的浓度。结果表明,该方法与酶联免疫吸附测定法之间没有明显差异,表明所建议方法具有良好的准确性和可用性。