Chavan Dimple, Chen Hui, Crum Mary, Vu Binh, Safari Mohammad, Smith Maxwell, Vekilov Peter, Conrad Jacinta C, Kourentzi Katerina, Willson Richard C
Department of Biology and Biochemistry, University of Houston, Houston, Texas 77204, USA.
Analyst. 2020 Jul 21;145(14):4942-4949. doi: 10.1039/d0an00134a. Epub 2020 Jun 5.
We have developed an immuno-PCR based diagnostic platform which couples detection antibodies to self-assembled, ultra-detectable DNA-avidin nanoparticles stabilized with poly(ethylene glycol) to link DNA amplification to target protein concentration. Electrostatic neutralization and cloaking of the PCR-amplifiable DNA labels by avidin and PEG coating reduces non-specific "stickiness" and enhances assay sensitivity. We further optimized the detectability of the nanoparticles by incorporating four repeats of a unique synthetic DNA PCR target into each nanoparticle. Using human chorionic gonadotropin hormone (hCG) as a model analyte, this platform was able to quantitate the target hCG protein in femtomolar concentrations using only standard laboratory equipment.
我们开发了一种基于免疫聚合酶链反应的诊断平台，该平台将检测抗体与通过聚乙二醇稳定的自组装、超可检测的DNA-抗生物素蛋白纳米颗粒相结合，以将DNA扩增与目标蛋白浓度联系起来。抗生物素蛋白和聚乙二醇涂层对可进行PCR扩增的DNA标签进行静电中和及包覆，减少了非特异性“粘性”并提高了检测灵敏度。我们通过在每个纳米颗粒中加入四个独特合成DNA PCR靶标的重复序列，进一步优化了纳米颗粒的可检测性。以人绒毛膜促性腺激素（hCG）作为模型分析物，该平台仅使用标准实验室设备就能定量飞摩尔浓度的目标hCG蛋白。
