Bridging the Gap in the Development of Gold Nanoparticle-based Enzyme Biosensing

缩小基于金纳米颗粒的酶生物传感的发展差距

• 批准号: EP/J013331/1
• 负责人: Molly Stevens
• 金额: $ 14.77万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FJ013331%2F1
• 关键词: Bridging; Gap; Development; Gold; Nanoparticle

Quantitatively measuring enzyme activity is a great challenge since the product of this activity is often altered by only a single molecular bond. Many current methods rely on the creation of fluorescent analogues that exhibit an increase or decrease in brightness when modified by enzymes. This approach is problematic, though, because the addition of a bulky fluorescent group can change the molecular interaction. Furthermore, it is often a slow and tedious process to redesign these tests for new applications. In contrast, we have developed an array of assays based on the unique optical properties of gold nanoparticles, which do not suffer from these drawbacks.By selectively functionalising the surface of gold nanoparticles, we are able to make them either aggregate or disperse in response to enzyme activation. This change in physical state then manifests itself as a dramatic colour change of the solution. This property naturally lends itself to both qualitative and quantitative measurements of enzyme activity. Over the past several years, we have developed and tested a number of assays based on this basic concept. The two most successful to date have been a protease assay and a lipase assay, both of which are of great clinical relevance. We have explored using the protease assay to test for HIV biomarkers and, since proteases are involved in a number of viral and parasitic infections, we expect that there will be many more applications to come. Lipases are also medically important, as their level in the body can become too high due to arthritis, acute sepsis and certain types of cancer. Because of this wide-ranging involvement, efforts are currently underway in the pharmaceutical industry to develop drugs that will target lipases.One of the great advantages of our technology is the fact that, in addition to diagnosing disease, it can also potentially help in the search for new drugs to treat those diseases. In contrast to methods that simply measure enzyme concentration, our method actually measures enzyme activity. We are just now starting to take advantage of this capability by using our assays to help screen chemical libraries for new drug candidates.In the course of this Follow-on fund project, we will address some of the practical questions involved in the transition from benchtop to bedside. In particular, we will look at scalability, manufacturing and storage and determine the commercial landscape and route forwards. These are concerns that are typically not addressed at the level of fundamental research but are important for the creation of a practical product. At the conclusion of this work, we will be in a strong position to immediately commercialise this technology, bringing its benefits to the broader healthcare market.

定量测量酶的活性是一个巨大的挑战，因为这种活性的产物往往只被单个分子键改变。许多当前的方法依赖于荧光类似物的创造，这些类似物在被酶修饰时表现出亮度的增加或减少。然而，这种方法是有问题的，因为添加一个庞大的荧光基团会改变分子的相互作用。此外，为新应用程序重新设计这些测试通常是一个缓慢而乏味的过程。相比之下，我们已经开发了一系列基于金纳米颗粒独特的光学特性的检测方法，这些方法没有这些缺点。通过选择性地功能化金纳米颗粒的表面，我们能够使它们在酶的激活下聚集或分散。这种物理状态的变化随后表现为溶液的剧烈颜色变化。这一特性自然地使酶活性的定性和定量测量成为可能。在过去的几年中，我们基于这一基本概念开发和测试了许多检测方法。迄今为止最成功的两种方法是蛋白酶测定法和脂肪酶测定法，这两种方法都具有很大的临床相关性。我们已经探索了使用蛋白酶测定来检测HIV生物标志物，由于蛋白酶与许多病毒和寄生虫感染有关，我们预计会有更多的应用。脂肪酶在医学上也很重要，因为它们在体内的水平可能因关节炎、急性败血症和某些类型的癌症而过高。由于这种广泛的参与，制药行业目前正在努力开发针对脂肪酶的药物。我们的技术的一大优势是，除了诊断疾病之外，它还可以潜在地帮助寻找治疗这些疾病的新药。与简单测量酶浓度的方法相比，我们的方法实际上是测量酶的活性。我们现在刚刚开始利用这种能力，通过使用我们的分析来帮助筛选新的候选药物的化学文库。在这个后续基金项目的过程中，我们将解决从台式到床边过渡中涉及的一些实际问题。特别是，我们将关注可扩展性、制造和存储，并确定商业前景和前进路线。这些问题通常不会在基础研究层面得到解决，但对于创建实用产品非常重要。在这项工作结束时，我们将处于一个强有力的位置，可以立即将这项技术商业化，将其好处带给更广泛的医疗保健市场。