Amplified Protein Detection and Identification through DNA-Conjugated M13 Bacteriophage
Amplified Protein Detection and Identification through DNA-Conjugated M13 Bacteriophage
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DOI：
10.1021/nn301565e
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发表时间：
2012-06-01
期刊：
影响因子：
17.1
通讯作者：
Cha, Jennifer N.
中科院分区：
文献类型：
--
作者：
Lee, Ju Hun;Domaille, Dylan W.;Cha, Jennifer N.
在临床和实验室环境中，对敏感的蛋白质检测和准确的鉴定仍然有很大的需求。为了使这些诊断具有临床价值，有必要开发具有高灵敏度但具有良好成本效益比的传感器。然而，这些传感平台中的许多都是热不稳定的，或者需要大量的材料合成、工程或制造。最近，我们证明了天然存在的M13噬菌体可以作为工程蛋白诊断的生物支架。这些病毒有5个丸状蛋白的拷贝，可以特异性地与目标抗原结合，还有数千个pVIII外壳蛋白，可以通过基因或化学修饰，与产生信号的物质发生反应，比如等离子体移动金纳米粒子（Au NPs）。在本报告中，我们发现dna偶联的M13噬菌体可以作为廉价的蛋白质传感器，在靶蛋白存在的情况下快速诱导颜色变化，同时也提供了在单独步骤中识别被检测抗原的能力。将特定DNA寡核苷酸的许多拷贝附加到每种病毒上，以创建可以与DNA偶联金纳米粒子杂交的噬菌体-DNA偶联物。在比色法阳性结果的情况下，抗原的身份也可以很容易地确定使用DNA微阵列。这节省了宝贵的资源，建立了一种快速、定量的方法，首先筛选抗原的存在，然后在必要时进行高度特异性的分型试验。
Sensitive protein detection and accurate identification continues to be in great demand for disease screening In clinical and laboratory settings. For these diagnostics to be of clinical value, it is necessary to develop sensors that have high sensitivity but favorable cost-to-benefit ratios. However, many of these sensing platforms are thermally unstable or require significant materials synthesis, engineering, or fabrication. Recently, we demonstrated that naturally occurring M13 bacteriophage can serve as biological scaffolds for engineering protein diagnostics. These viruses have five copies of the pill protein, which can bind specifically to target antigens, and thousands of pVIII coat proteins, which can be genetically or chemically modified to react with signal-producing materials, such as plasmon-shifting gold nanoparticles (Au NPs). In this report, we show that DNA-conjugated M13 bacteriophage can act as inexpensive protein sensors that can rapidly induce a color change in the presence of a target protein yet also offer the ability to identify the detected antigen in a separate step. Many copies of a specific DNA oligonucleotide were appended to each virus to create phage-DNA conjugates that can hybridize with DNA-conjugated gold nanoparticles. In the case of a colorimetric positive result, the identity of the antigen can also be easily determined by using a DNA microarray. This saves precious resources by establishing a rapid, quantitative method to first screen for the presence of antigen followed by a highly specific typing assay if necessary.