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用于灵敏和选择性检测碱性磷酸酶的 VS 量子点无标记荧光探针。

VS quantum dot label-free fluorescent probe for sensitive and selective detection of ALP.

机构信息

Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun, Jilin, 130012, China.

出版信息

Anal Bioanal Chem. 2018 Feb;410(5):1417-1426. doi: 10.1007/s00216-017-0778-8. Epub 2017 Dec 18.

Abstract

As an essential enzyme highly associated with various human diseases, alkaline phosphatase (ALP) plays an important role in human tissues. Developing new materials and strategies for monitoring ALP is thus important. We have developed a novel label-free fluorescent sensing system for ALP activity that is based on the "turn-on" fluorescence of VS quantum dots. The fluorescence of VS quantum dots quenched by Fe can be restored by ascorbic acid, which is generated by hydrolysis of L-ascorbic acid 2-phosphate catalyzed by ALP. Rapid, convenient, and sensitive detection of ALP is achieved in the range from 3 to 1000 U/L (R =0.9985), with a detection limit of 0.27 U/L. The proposed sensor exhibits excellent selectivity for ALP compared with other enzymes and proteins, such as glucose oxidase, lysozyme, trypsin, human serum albumin, and bovine serum albumin. The reliability for ALP determination in human serum plasma has been demonstrated with satisfactory recovery, revealing promising application in clinical diagnosis and biomedical research. Graphical abstract Hydrothermally synthesized VS quantum dots serving as a novel turn-on fluorescent probe for detection of alkaline phosphatase (ALP) activity. AA L-ascorbic acid, AAP L-ascorbic acid 2-phosphate, NAC N-acetyl-L-cysteine.

摘要

作为一种与多种人类疾病密切相关的重要酶,碱性磷酸酶(ALP)在人体组织中发挥着重要作用。因此,开发用于监测 ALP 的新材料和策略非常重要。我们已经开发出一种基于 VS 量子点的“开启”荧光的新型无标记荧光传感系统,用于 ALP 活性检测。VS 量子点的荧光可以被 Fe 猝灭,但可以被 ALP 催化水解 L-抗坏血酸 2-磷酸产生的抗坏血酸(AA)所恢复。在 3 至 1000 U/L(R =0.9985)的范围内,实现了对 ALP 的快速、方便和灵敏检测,检测限为 0.27 U/L。与其他酶和蛋白质(如葡萄糖氧化酶、溶菌酶、胰蛋白酶、人血清白蛋白和牛血清白蛋白)相比,该传感器对 ALP 具有优异的选择性。在人血清血浆中进行 ALP 测定的可靠性已得到证明,具有令人满意的回收率,有望在临床诊断和生物医学研究中得到应用。

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