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Digital counting of single semiconducting polymer nanoparticles for the detection of alkaline phosphatase
Nanoscale ( IF 5.8 ) Pub Date : 2021-2-1 , DOI: 10.1039/d0nr09232k Shumin Wang 1, 2, 3, 4, 5 , Mengna Huang 1, 2, 3, 4, 5 , Jianhao Hua 6, 7, 8, 9, 10 , Lin Wei 1, 2, 3, 4, 5 , Shen Lin 6, 7, 8, 9, 10 , Lehui Xiao 6, 7, 8, 9, 10
Nanoscale ( IF 5.8 ) Pub Date : 2021-2-1 , DOI: 10.1039/d0nr09232k Shumin Wang 1, 2, 3, 4, 5 , Mengna Huang 1, 2, 3, 4, 5 , Jianhao Hua 6, 7, 8, 9, 10 , Lin Wei 1, 2, 3, 4, 5 , Shen Lin 6, 7, 8, 9, 10 , Lehui Xiao 6, 7, 8, 9, 10
Affiliation
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Alkaline phosphatase (ALP) as a necessary hydrolase in phosphate metabolism is closely related to various diseases. Ultrasensitive detection of ALP with a convenient and sensitive method is of fundamental importance. In this work, a fluorescence resonance energy transfer (FRET)-based single-particle enumeration (SPE) method is proposed for the quantitative analysis of ALP. This strategy is based on the effective fluorescence suppression by a polydopamine (PDA) shell on the surface of semiconducting polymer nanoparticles (SPNs). PDA with broadband absorption in the UV-vis region can serve as an excellent quencher for SPNs. However, ascorbic acid (AA), the product of the hydrolysis of 2-phosphate-L-ascorbic acid trisodium salt (AAP) in the presence of ALP, can effectively inhibit the self-polymerization of dopamine (DA) to form a PDA layer. Therefore, ALP can be accurately quantified by counting the concentration-related fluorescent particles in the fluorescence image. A linear range from 0.031 to 12.4 μU mL−1 and a limit-of-detection (LOD) of 0.01 μU mL−1 for ALP determination are achieved. The spiked recoveries for ALP determination in a human serum sample are between 90% and 108% with RSD less than 3.1%. In summary, this convenient and sensitive approach proposed here provides promising prospects for ALP detection in a complex biological matrix.
中文翻译：
用于检测碱性磷酸酶的单个半导体聚合物纳米粒子的数字计数
碱性磷酸酶（ALP）是磷酸盐代谢中必需的水解酶，与多种疾病密切相关。用方便和灵敏的方法对ALP进行超灵敏检测至关重要。在这项工作中，提出了一种基于荧光共振能量转移（FRET）的单颗粒枚举（SPE）方法用于ALP定量分析的方法。该策略基于半导体聚合物纳米粒子（SPN）表面上的聚多巴胺（PDA）外壳对荧光的有效抑制。在紫外可见区域具有宽带吸收能力的PDA可以用作SPN的出色淬灭剂。但是，抗坏血酸（AA）是2-磷酸-L的水解产物-抗坏血酸三钠盐（AAP）在ALP的存在下，可有效抑制多巴胺（DA）的自聚合形成PDA层。因此，通过对荧光图像中的浓度相关的荧光粒子进行计数，可以准确地定量ALP。的线性范围为0.031至12.4μU毫升-1 0.01μU毫升和一个极限的检测（LOD）-1为ALP测定来实现的。用于测定人血清样品中ALP的加标回收率在90％至108％之间，RSD小于3.1％。总之，这里提出的这种方便而灵敏的方法为在复杂的生物基质中进行ALP检测提供了有希望的前景。
更新日期：2021-02-25
中文翻译：
用于检测碱性磷酸酶的单个半导体聚合物纳米粒子的数字计数
碱性磷酸酶（ALP）是磷酸盐代谢中必需的水解酶，与多种疾病密切相关。用方便和灵敏的方法对ALP进行超灵敏检测至关重要。在这项工作中，提出了一种基于荧光共振能量转移（FRET）的单颗粒枚举（SPE）方法用于ALP定量分析的方法。该策略基于半导体聚合物纳米粒子（SPN）表面上的聚多巴胺（PDA）外壳对荧光的有效抑制。在紫外可见区域具有宽带吸收能力的PDA可以用作SPN的出色淬灭剂。但是，抗坏血酸（AA）是2-磷酸-L的水解产物-抗坏血酸三钠盐（AAP）在ALP的存在下，可有效抑制多巴胺（DA）的自聚合形成PDA层。因此，通过对荧光图像中的浓度相关的荧光粒子进行计数，可以准确地定量ALP。的线性范围为0.031至12.4μU毫升-1 0.01μU毫升和一个极限的检测（LOD）-1为ALP测定来实现的。用于测定人血清样品中ALP的加标回收率在90％至108％之间，RSD小于3.1％。总之，这里提出的这种方便而灵敏的方法为在复杂的生物基质中进行ALP检测提供了有希望的前景。
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