背景:作为黑素细胞病变的重要生物标志物,酪氨酸酶(TYR)在黑色素相关疾病的临床诊断和治疗中起着至关重要的作用。因此,重要的是开发稳健的方法来评估TYR活性。共价有机骨架(COFs)因其独特的性质而备受关注,包括高化学稳定性,良好的生物相容性,与有机染料相比,表面积大,贵金属纳米团簇,和半导体量子点。然而,大多数COF不溶于水并且表现出弱的荧光发射或没有荧光发射。因此,仍然高度期望开发用于检测生物样品中的TYR活性的水溶性荧光COF。
结果:在这项工作中,建立了一种灵敏,简便的基于荧光COF的荧光法,用于检测人血清样品中的TYR活性。水溶性COF是通过4'的缩聚制备的,4,4\'\'\'\'\'\',4\'\'\'\'\'\'\'\''-(1,2-亚乙基)四[1,1'-联苯]-4-甲醛和2,4,6-三-(4-氨基苯基)-三嗪。所得COF显示黄绿色荧光,在560nm处具有最大发射峰。酪氨酸被TYR催化以产生黑色素样聚合物,其在COF的表面上形成涂层并且由于荧光共振能量转移而有效地猝灭其荧光。所提出的方法在0.5-80U/L范围内表现出很强的线性相关性,检测限为0.09U/L。此外,曲酸的检测限,作为代表性的TYR抑制剂,测定为0.0004μg/mL。
结论:我们提出的荧光传感平台具有出色的选择性,灵敏度,和令人满意的人血清样品的回收率,这对于黑色素相关疾病的临床诊断至关重要。此外,所提出的方法进一步用于筛选TYR抑制剂,提示在临床治疗和药物研究中的潜在应用。
BACKGROUND: 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.
RESULTS: 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.
CONCLUSIONS: 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.