Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, CPZN 5206, Boston, Massachusetts 02114, USA.
ACS Nano. 2011 Apr 26;5(4):3204-13. doi: 10.1021/nn200333m. Epub 2011 Mar 8.
Nanomaterials offer unique physical properties that make them ideal biosensors for scant cell populations. However, specific targeting of nanoparticles to intracellular proteins has been challenging. Here, we describe a technique to improve intracellular biomarker sensing using nanoparticles that is based on bioorthogonal chemistry. Using trans-cyclooctene-modified affinity ligands that are administered to semipermeabilized cells and revealed by cycloaddition reaction with tetrazine-conjugated nanoparticles, we demonstrate site-specific amplification of nanomaterial binding. We also show that this technique is capable of sensing protein biomarkers and phosho-protein signal mediators, both within the cytosol and nucleus, via magnetic or fluorescent modalities. We expect the described method will have broad applications in nanomaterial-based diagnostics and therapeutics.
纳米材料具有独特的物理性质，使其成为检测稀有细胞群体的理想生物传感器。然而，将纳米颗粒特异性靶向细胞内蛋白质一直具有挑战性。在这里，我们描述了一种使用基于生物正交化学的纳米颗粒来提高细胞内生物标志物检测的技术。我们使用顺式环辛烯修饰的亲和配体，将其施用于半透化细胞中，并通过与四嗪缀合的纳米颗粒的环加成反应进行揭示，从而证明了纳米材料结合的特异性扩增。我们还表明，该技术能够通过磁性或荧光模式在细胞质和核内检测蛋白质生物标志物和磷酸化蛋白信号介质。我们预计所描述的方法将在基于纳米材料的诊断和治疗中具有广泛的应用。
