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Abstract：
The effect of preformed protein coronas on immunoassays for Dengue nonstructural protein 1 (NS1) immunoassays was investigated. The composition of the protein corona that forms around nanoparticle-antibody conjugates in human serum was characterized, and selected proteins from the corona were used for preformed coronas (human serum albumin and apolipoprotein A1). Coronas were formed and characterized by dynamic light scattering (DLS), and the nanoparticle-conjugate was probed by optical absorption spectroscopy. Immunoassays were run, and performance was quantified by analyzing the strip intensity as a function of NS1 concentration. The preformed coronas influenced the limit of detection (LOD) of the assay and the affinity for the NS1 target (KD). The resulting KD and LODs for the NP-Ab-ApoA1 immunoprobes were 0.83 nM and 1.24 nM, respectively. For the NP-Ab -HSA coronas, the test line intensity was lower by 33% at a given NS1 concentration than for the NP-Ab immunoprobes, and KD was 0.14 nM, a slightly higher affinity. Due to the relatively large error of the negative control, a meaningful LOD for the NP-Ab with HSA coronas could not be determined.
摘要：
研究了预制蛋白电晕对登革热非结构蛋白1（NS1）免疫测定的影响。表征了人血清中纳米颗粒-抗体缀合物周围形成的蛋白质冠的组成，并且从电晕中选择的蛋白质用于预先形成的冠状蛋白（人血清白蛋白和载脂蛋白A1）。形成日冕，并通过动态光散射（DLS）进行表征，并通过光学吸收光谱法探测纳米颗粒-缀合物。进行了免疫测定，并且通过分析作为NS1浓度的函数的条带强度来量化性能。预先形成的电晕会影响测定的检测限（LOD）和对NS1靶标（KD）的亲和力。所得NP-Ab-ApoA1免疫探针的KD和LOD为0.83nM和1.24nM，分别。对于NP-Ab-HSA冠状病毒，在给定的NS1浓度下，测试线强度比NP-Ab免疫探针低33%，KD为0.14nM，稍高的亲和力。由于阴性对照误差较大,无法确定NP-Ab合并HSA冠状蛋白的有意义的LOD.
