Department of Chemistry, The University of Texas at Austin, 100 E. 24th Street, Stop A1590, Austin, Texas 78712-1224, United States.
Interactives Executive Excellence LLC, Austin, Texas 78733, United States.
ACS Sens. 2021 May 28;6(5):1956-1962. doi: 10.1021/acssensors.1c00446. Epub 2021 Apr 22.
The effect of serum on electrochemical detection of bioassays having silver nanoparticle (AgNP) detection labels was investigated. Both a model assay and an antigen-specific sandwich bioassay for the heart failure marker NT-proBNP were examined. In both cases, the AgNP labels were conjugated to a detection antibody. Electrochemical detection was carried out using a galvanic exchange/anodic stripping voltammetry method in which Au exchanges with AgNP labels. The assays were carried out using a paper-based electrode platform. The bioassays were exposed to different serum conditions prior to and during detection. There are three important outcomes reported in this article. First, both the model- and antigen-specific assays could be formed in undiluted serum with no detectable interferences from the serum components. Second, to achieve the maximum possible electrochemical signal, the highest percentage of serum that can remain in an assay buffer during electrochemical detection is 0.25% when no washing is performed. The assay results are rendered inaccurate when 0.50% or more of serum is present. Third, the factors inhibiting galvanic exchange in serum probably relate to surface adsorption of biomolecules onto the AgNP labels, chelation of Au by serum components, or both. The results reported here provide general guidance for using metal NP labels for electrochemical assays in biofluids.
研究了血清对具有银纳米粒子(AgNP)检测标签的生物分析电化学检测的影响。对心力衰竭标志物 NT-proBNP 的模型分析和抗原特异性夹心生物分析都进行了检查。在这两种情况下,AgNP 标签都与检测抗体连接。电化学检测使用电化学生换/阳极溶出伏安法进行,其中 Au 与 AgNP 标签交换。分析使用基于纸张的电极平台进行。在检测之前和期间,生物分析暴露于不同的血清条件下。本文报道了三个重要结果。首先,即使在没有血清成分干扰的情况下,模型和抗原特异性分析都可以在未稀释的血清中形成。其次,为了获得最大可能的电化学信号,在不进行洗涤的情况下,在电化学检测过程中,分析缓冲液中保留的血清的最高百分比为 0.25%。当存在 0.50%或更多的血清时,分析结果会变得不准确。第三,血清中抑制电化学生换的因素可能与生物分子在 AgNP 标签上的表面吸附、血清成分对 Au 的螯合或两者都有关。这里报道的结果为在生物流体中使用金属 NP 标签进行电化学分析提供了一般性指导。