School of Science, Xihua University, Chengdu, 610039, China.
Department of Clinical Laboratory, Zigong First People's Hospital, Zigong, 643000, Sichuan, China.
Anal Chim Acta. 2024 Sep 1;1320:343026. doi: 10.1016/j.aca.2024.343026. Epub 2024 Jul 25.
As a significant biomarker of melanocytic lesions, tyrosinase (TYR) plays an essential role in the clinical diagnosis and treatment of melanin-related diseases. Thus, it is important to develop robust methods for assessing TYR activity. Covalent organic frameworks (COFs) have garnered considerable attention owing to their unique properties, including high chemical stability, good biocompatibility, and large surface area compared with organic dyes, noble metal nanoclusters, and semiconductor quantum dots. However, most COFs are insoluble in water and exhibit weak or no fluorescence emission. Therefore, the development of a water-soluble fluorescent COF for detecting TYR activity in biological samples remains highly desired.
In this work, a sensitive and facile fluorometric method based on fluorescent COF was constructed for the detection of TYR activity in human serum samples. The water-soluble COF was fabricated through the condensation polymerization of 4',4‴,4''''',4'''''''-(1,2-ethene-diylidene) tetrakis [1,1'-biphenyl]-4-carboxaldehyde and 2,4,6-tris-(4-aminophenyl)-triazine. The resulting COF displayed yellow-green fluorescence with a maximum emission peak at 560 nm. Tyrosine was catalyzed by TYR to produce melanin-like polymers which formed a coating on the surface of COF and effectively quenched its fluorescence due to fluorescence resonance energy transfer. The proposed approach demonstrated a strong linear correlation in the range of 0.5-80 U/L with a low detection limit of 0.09 U/L. Additionally, the limit of detection for kojic acid, serving as a representative TYR inhibitor, was determined to be 0.0004 μg/mL.
Our proposed fluorometric sensing platform exhibited exceptional selectivity, sensitivity, and satisfactory recoveries in human serum samples, which is of paramount importance for the clinical diagnostics of melanin-related diseases. Furthermore, the proposed approach was further employed for the screening of TYR inhibitors, suggesting the potential applications in clinical treatment and pharmaceutical research.
酪氨酸酶（TYR）作为黑素细胞病变的重要生物标志物，在黑色素相关疾病的临床诊断和治疗中发挥着重要作用。因此，开发用于评估 TYR 活性的稳健方法非常重要。与有机染料、贵金属纳米团簇和半导体量子点相比，共价有机框架（COFs）具有独特的性质，包括高化学稳定性、良好的生物相容性和大的表面积，因此备受关注。然而，大多数 COFs 不溶于水，并且表现出弱的或没有荧光发射。因此，开发用于检测生物样品中 TYR 活性的水溶性荧光 COF 仍然是非常需要的。
在这项工作中，构建了一种基于荧光 COF 的灵敏、简便的荧光法，用于检测人血清样品中的 TYR 活性。通过 4'，4"，4'''，4''''''''-（1,2-亚乙基二基）四[1,1'-联苯]-4-甲醛和 2,4,6-三-（4-氨基苯基）-均三嗪的缩聚反应制备了水溶性 COF。所得 COF 显示出黄绿色荧光，最大发射峰位于 560nm。酪氨酸被 TYR 催化生成黑色素样聚合物，这些聚合物在 COF 表面形成涂层，由于荧光共振能量转移，有效地猝灭了其荧光。所提出的方法在 0.5-80 U/L 的范围内表现出很强的线性相关性，检测限低至 0.09 U/L。此外，作为代表性的 TYR 抑制剂的曲酸的检测限被确定为 0.0004μg/mL。
我们提出的荧光传感平台在人血清样品中表现出优异的选择性、灵敏度和令人满意的回收率，这对黑色素相关疾病的临床诊断至关重要。此外，该方法还进一步用于 TYR 抑制剂的筛选，表明其在临床治疗和药物研究方面具有潜在的应用。
