Abstract

The discovery and application of analyte-triggered fluorophore generation or fluorogenic reaction are significant and beneficial to the development of novel fluorescence (FL) analysis method. In this study, for the first time, we have reported a fluorogenic reaction to prepare fluorescent silicon-containing polymer dots (Si-PDs) by simply mixing N-[3-(trimethoxysilyl)propyl]ethylenediamine (DAMO) and hydroquinone (HQ) in aqueous solution at ambient temperature. Inspired by the alkaline phosphatase (ALP)-catalyzed hydrolysis of the substrate sodium 4-hydroxyphenyl phosphate (4-HPP) into HQ and the resultant HQ-controlled intense green Si-PDs generation, we have established a straightforward ALP activity assay by innovatively employing commercially available 4-HPP as the substrate. More significantly, the specific preparation method, clear formation mechanism and excellent performance enable the Si-PDs as well as its generation process to develop facile and attractive FL immunoassay. With the help of the universal ALP-based enzyme-linked immunosorbent assay (ELISA) platform and corresponding antibody, a convenient and conceptual ALP-based fluorescent ELISA has been constructed and applied in sensing cardiac troponin I (cTnI), a well-known biomarker of acute myocardial infarction. Our research via in situ formation of fluorescent nanomaterials has great potential application in ALP activity assay, inhibitor screening, and disease diagnosis.

中文翻译：

---

用于碱性磷酸酶活性检测和免疫测定的荧光含硅聚合物点的原位形成

摘要

被分析物触发的荧光团产生或荧光反应的发现和应用具有重要意义，并有益于新型荧光分析方法的发展。在这项研究中，我们首次报道了一种荧光反应，该反应通过简单地混合N来制备含荧光硅的聚合物点（Si-PDs）-[3-（三甲氧基甲硅烷基）丙基]乙二胺（DAMO）和对苯二酚（HQ）在室温下的水溶液中。受碱性磷酸酶（ALP）催化的底物4-羟基苯基磷酸钠（4-HPP）水解成HQ以及由此产生的由HQ控制的深绿色Si-PDs产生的启发，我们通过创新地采用了直接建立ALP活性的方法市售的4-HPP作为底物。更重要的是，特定的制备方法，清晰的形成机理和出色的性能使Si-PDs及其生成过程得以发展出简便而诱人的FL免疫测定法。借助基于通用ALP的酶联免疫吸附测定（ELISA）平台和相应的抗体，已经构建了一种方便且基于概念的基于ALP的荧光ELISA，并将其用于检测心肌肌钙蛋白I（cTnI），这是急性心肌梗死的著名生物标志物。我们的研究通过荧光纳米材料的原位形成在ALP活性测定，抑制剂筛选和疾病诊断中具有巨大的潜力。