Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, Chongqing, 400715, PR China.
Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, Chongqing, 400715, PR China.
Anal Biochem. 2019 Jul 15;577:82-88. doi: 10.1016/j.ab.2019.04.020. Epub 2019 Apr 26.
Biothiols play critical roles in many biological processes and their aberrant is related to a variety of syndromes. A simple and reliable colorimetric method is developed in this work for biothiols detection based on an oxidase mimic, a metal organic framework (MOF) MIL-53(Fe), and a peroxidase substrate 3,3',5,5'-tetramethylbenzidine (TMB). In this design, MIL-53(Fe) is utilized to catalyze the conversion of TMB to a blue colored 3,3',5,5'-tetramethylbenzidine diimine, which can be read on a spectrophotometer at 652 nm. The oxidation-induced blue color generation can be efficiently inhibited by biothiols, thus a colorimetric analytical method is proposed for biothiols detection based on the above system. Under optimal conditions, a linear relationship in a range from 1 to 100 μM and a limit of detection (LOD) at 120 nM are achieved with Cys as a model target. The developed platform is further applied to evaluate cellular biothiols in normal (RWPE-1) and cancer (LNCap) cell lines, revealing that the overall biothiols level in LNCap is much higher than that in RWPE-1. This work renders a powerful tool for identifying cancer cells in a simple manner for biomedical diagnosis associated with biothiols.
生物硫醇在许多生物过程中发挥着关键作用,它们的异常与多种综合征有关。本工作基于一种氧化酶模拟物、一种金属有机骨架(MOF)MIL-53(Fe)和一种过氧化物酶底物 3,3',5,5'-四甲基联苯胺(TMB),开发了一种用于生物硫醇检测的简单可靠的比色法。在该设计中,MIL-53(Fe) 用于催化 TMB 转化为蓝色的 3,3',5,5'-四甲基联苯二亚胺,可在分光光度计上于 652nm 处读取。生物硫醇可有效抑制氧化诱导的蓝色生成,因此基于上述体系提出了一种用于生物硫醇检测的比色分析方法。在最佳条件下,以半胱氨酸为模型目标,检测范围为 1 至 100µM,检测限(LOD)为 120nM。该平台进一步用于评估正常(RWPE-1)和癌症(LNCap)细胞系中的细胞生物硫醇,结果表明 LNCap 中的总生物硫醇水平明显高于 RWPE-1。这项工作为生物硫醇相关的生物医学诊断中以简单的方式识别癌细胞提供了有力的工具。