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基于单磷酸腺苷保护的双金属纳米簇的间日疟原虫乳酸脱氢酶新型荧光探针。

A novel fluorescence probe of Plasmodium vivax lactate dehydrogenase based on adenosine monophosphate protected bimetallic nanoclusters.

机构信息

State Key Laboratory of Supramolecular Structure and Materials, Institute of Theoretical Chemistry, Jilin University, No. 2699 Qianjin Street, Changchun, 130012, China.

School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China.

出版信息

Talanta. 2020 Jun 1;213:120850. doi: 10.1016/j.talanta.2020.120850. Epub 2020 Feb 14.

Abstract

Specific detection of Plasmodium vivax lactate dehydrogenase (PvLDH), an important biomarker of malaria, remains a significant challenge. Herein, adenosine monophosphate protected gold-silver bimetallic nanoclusters, Au-AgNCs@AMP were used as a specific and sensitive fluorescence probe to detect PvLDH. After optimizing, a linear response was shown over a wide concentration range (10-100 nM) and an extremely low limit of detection (LOD) at 0.10 nM (3.7 ng mL) was achieved finally. Albeit the method was able to detect PvLDH sensitively, it could not discriminate different types of LDHs. Consequently, Al was employed as an "assistant agent", which induced an assay capacity to discriminate PvLDH from other LDHs. The bimetallic nanoclusters inhibited the activity of PvLDH, suggesting it bound near the active site of PvLDH with high affinity. Zeta potential and UV-vis absorption experiments showed that electrostatic interaction was the main driving force for the interaction between the nanoclusters and PvLDH. Through chemical modification it indicated free thiol groups in PvLDH played an implant role in the interaction. Overall, the fluorescence enhancement and blue-shift were attributed to assembly-induced emission enhancement (AIEE) and hydrophobic transfer. The present study provides a simple, robust, and easy-to-perform approach to detect PvLDH with high sensitivity and selectivity, with significant potential for malaria diagnosis in the developing world.

摘要

特异性检测间日疟原虫乳酸脱氢酶(PvLDH),这是疟疾的一个重要生物标志物,仍然是一个重大挑战。在此,我们使用腺嘌呤单核苷酸保护的金-银双金属纳米团簇,Au-AgNCs@AMP 作为一种特异性和灵敏的荧光探针来检测 PvLDH。经过优化,该方法在较宽的浓度范围内(10-100 nM)呈现线性响应,并且最终达到极低的检测限(LOD)为 0.10 nM(3.7 ng mL)。虽然该方法能够灵敏地检测 PvLDH,但它不能区分不同类型的 LDH。因此,我们使用 Al 作为“辅助剂”,使其具有区分 PvLDH 与其他 LDH 的能力。双金属纳米团簇抑制了 PvLDH 的活性,表明它与 PvLDH 的活性位点结合具有高亲和力。Zeta 电位和紫外-可见吸收实验表明,静电相互作用是纳米团簇与 PvLDH 相互作用的主要驱动力。通过化学修饰表明 PvLDH 中的游离巯基在相互作用中起重要作用。总的来说,荧光增强和蓝移归因于组装诱导的发射增强(AIEE)和疏水性转移。本研究提供了一种简单、稳健、易于操作的方法,用于高灵敏度和选择性地检测 PvLDH,具有在发展中国家进行疟疾诊断的巨大潜力。

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