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Localized Visualization and Autonomous Detection of Cell Surface Receptor Clusters Using DNA Proximity Circuit

机译:使用DNA接近电路的局部可视化和自主检测细胞表面受体簇

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摘要

Cell surface receptors play an important role in mediating cell communication and are used as disease biomarkers and therapeutic targets. We present a one-pot molecular toolbox, which we term the split proximity circuit (SPC), for the autonomous detection and visualization of cell surface receptor clusters. Detection was powered by antibody recognition and a series of autonomous DNA hybridization to achieve localized, enzyme-free signal amplification. The system under study was the human epidermal growth factor receptor (HER) family, that is, HER2:HER2 homodimer and HER2:HER3 heterodimer, both in cell lysate and in situ on fixed whole cells. The detection and imaging of receptors were carried out using standard microplate scans and confocal microscopy, respectively. The circuit operated specifically with minimal leakages and successfully captured the receptor expression profiles on three cell types without any intermediate washing steps.
机译:细胞表面受体在介导细胞通信中起重要作用,并用作疾病生物标志物和治疗靶标。 我们介绍了一个单壶分子工具箱,我们术语术语分离接近电路(SPC),用于细胞表面受体簇的自主检测和可视化。 检测由抗体识别和一系列自主DNA杂交提供动力,以实现局部化的无酶信号放大。 正在研究的系统是人体表皮生长因子受体(她的)家族,即HER2:HER2:HER2同源二聚体和HER2:HER3异二聚体,在细胞裂解物和原位上固定的整个细胞。 使用标准微孔板扫描和共聚焦显微镜分别进行受体的检测和成像。 电路专门操作,具有最小的泄漏,并在没有任何中间洗涤步骤的情况下成功地捕获了三种细胞类型的受体表达谱。

著录项

  • 来源
    《Analytical chemistry》 |2018年第10期|共6页
  • 作者单位

    Department of Chemical and Biomolecular Engineering National University of Singapore Singapore 117585 Singapore;

    Department of Chemical and Biomolecular Engineering National University of Singapore Singapore 117585 Singapore;

    Department of Anatomy Yong Loo Lin School of Medicine National University of Singapore Singapore 117594 Singapore;

    Department of Anatomy Yong Loo Lin School of Medicine National University of Singapore Singapore 117594 Singapore;

    Department of Anatomy Yong Loo Lin School of Medicine National University of Singapore Singapore 117594 Singapore;

    Department of Chemical and Biomolecular Engineering National University of Singapore Singapore 117585 Singapore;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
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