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通过原子转移自由基聚合对碱性磷酸酶活性进行电化学检测。

Electrochemical detection of alkaline phosphatase activity via atom transfer radical polymerization.

作者信息

Li Xiaofei, Lu Jing, Li Zutian, Yang Haoyuan, Li Weiming, Liu Yanju, Miao Mingsan

机构信息

Pharmacy College, Henan University of Chinese Medicine, Zhengzhou 450046, PR China.

The Third Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450003, PR China.

出版信息

Bioelectrochemistry. 2022 Apr;144:107998. doi: 10.1016/j.bioelechem.2021.107998. Epub 2021 Nov 16.

Abstract

Alkaline phosphatase (ALP) activity is a diagnostic indicator for a variety of clinical diseases. In this study, an electrochemical method for detecting ALP activity through activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) was developed. Specifically, 3-mercaptopropionic (MPA) was firstly fixed on the electrode through sulfur-gold bonding. Subsequently, α-bromophenylacetic acid (BPAA) as initiator was attached to MPA through the recognized carboxylate-Zr-phosphate chemistry. Finally, in the existence of ALP, L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate (AAPS) was hydrolyzed to produce ascorbic acid (AA) which participated in the ARGET ATRP reaction, grafting polymer containing plenty of ferrocene electroactive probes on the surface of electrode. Under optimal experimental conditions, this method had a linear scope of 20-200 mU mL, and a limit of detection (LOD) of 1.64 mU mL. In addition, the proposed method had good selectivity as well as anti-interference capability, with satisfactory results in inhibition rate and human serum experiments. By merits of good analytical performance, easy operation, and low cost, such a method for ALP activity detection has promising applications in ALP-related disease detection and inhibitor screening.

摘要

碱性磷酸酶(ALP)活性是多种临床疾病的诊断指标。在本研究中,开发了一种通过电子转移原子转移自由基聚合(ARGET ATRP)再生的活化剂来检测ALP活性的电化学方法。具体而言,首先通过硫-金键合将3-巯基丙酸(MPA)固定在电极上。随后,作为引发剂的α-溴苯乙酸(BPAA)通过公认的羧酸盐-锆-磷酸盐化学方法连接到MPA上。最后,在碱性磷酸酶存在的情况下,L-抗坏血酸2-磷酸倍半镁盐水合物(AAPS)水解生成抗坏血酸(AA),其参与ARGET ATRP反应,在电极表面接枝含有大量二茂铁电活性探针的聚合物。在最佳实验条件下,该方法的线性范围为20-200 mU mL,检测限(LOD)为1.64 mU mL。此外,该方法具有良好的选择性和抗干扰能力,在抑制率和人血清实验中均取得了满意的结果。由于具有良好的分析性能、操作简便和成本低廉等优点,这种检测碱性磷酸酶活性的方法在碱性磷酸酶相关疾病检测和抑制剂筛选中具有广阔的应用前景。

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