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An Enzyme Cascade-Triggered Fluorogenic and Chromogenic Reaction Applied in Enzyme Activity Assay and Immunoassay

机译:酶活性测定和免疫测定中施用酶级联触发的荧光和发色反应

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摘要

An enzyme cascade-triggered reaction with novel signal generation mechanism is beneficial for the development and insight of the enzyme cascade, which is extensively used for signal transduction in potential applications. Inspired by the fluorogenic and chromogenic reaction between dopamine and resorcinol, and the specific catalytic properties of alkaline phosphatase (ALP) and tyrosinase, we designed and synthesized an unconventional substrate of ALP, named p-aminoethyl-phenyl phosphate disodium salt (PAPP). As expected, the ALP and tyrosinase-incubated PAPP solution exhibited pale yellow with intense blue fluorescence upon addition of resorcinol, owing to the ALP-catalyzed transformation of PAPP into an intermediate tyramine, and the tyrosinase-catalyzed hydroxylation of tyramine to dopamine, as well as the specific reaction between dopamine and resorcinol. Therefore, an enzyme cascade system has been developed herein based on the ALP and tyrosinase coupled enzymes-triggered fluorogenic and chromogenic reaction. According to the direct relationship between the activity of ALP/tyrosinase and absorbance/fluorescence intensity of the resultant solution, the proposed enzyme cascade-triggered reaction was utilized for assaying ALP and tyrosinase activity with fluorometric and colorimetric dual-readout signals. Furthermore, such enzyme cascade catalysis process was integrated into the ALP-based cascade enzyme-linked immunosorbent assay with dual-readout signals, resulting in the sensitive detection of cardiac troponin I in diluted serum.
机译:与新型信号发电机制的酶级联触发反应有利于酶级联的开发和洞察力,其广泛用于潜在应用中的信号转导。通过多巴胺和间苯二酚之间的荧光和发色反应的启发,以及碱性磷酸酶(ALP)和酪氨酸酶的特定催化性能,我们设计和合成了ALP的非常规基材,称为对氨基乙基 - 磷酸二钠盐(PAPP)。正如预期的那样,由于PAPP到中间酪胺的Alp催化转化,并且酪氨酸酶催化剂的酪氨酸酶催化剂的酪氨酸酶催化剂的酪氨酸酶催化剂,并且酪蛋白溶解的浅黄色,并且在中间酪氨酸中,并且酪氨酸酶催化剂的酪氨酸酶催化剂的酪氨酸酶催化剂的溶血酶催化,以及多巴胺的酪氨酸酶催化剂,以及多巴胺的酪氨酸酶催化剂作为多巴胺和间苯二酚之间的特异性反应。因此,基于AlP和酪氨酸酶偶联酶触发的荧光和发色反应,已经在本文中开发了一种酶级联系统。根据所得溶液的ALP /酪氨酸酶和吸光度/荧光强度的活性之间的直接关系,利用所提出的酶级联触发反应进行荧光和比色双读信号测定AlP和酪氨酸酶活性。此外,将这种酶级联催化过程整合到基于ALP的级联酶联免疫吸附测定中,具有双读出信号,导致稀释血清中的心肌肌钙蛋白I的敏感性检测。

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  • 来源
    《Analytical chemistry》 |2018年第12期|共7页
  • 作者单位

    Chinese Acad Sci Changchun Inst Appl Chem State Key Lab Electroanalyt Chem Changchun 130022 Jilin Peoples R China;

    Chinese Acad Sci Changchun Inst Appl Chem State Key Lab Electroanalyt Chem Changchun 130022 Jilin Peoples R China;

    Chinese Acad Sci Changchun Inst Appl Chem State Key Lab Electroanalyt Chem Changchun 130022 Jilin Peoples R China;

    Chinese Acad Sci Changchun Inst Appl Chem State Key Lab Electroanalyt Chem Changchun 130022 Jilin Peoples R China;

    Chinese Acad Sci Changchun Inst Appl Chem State Key Lab Electroanalyt Chem Changchun 130022 Jilin Peoples R China;

    Chinese Acad Sci Changchun Inst Appl Chem State Key Lab Electroanalyt Chem Changchun 130022 Jilin Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
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