Self-regenerating, suspended-phase whole-cell biosensor system employing micro-chemostat and cell engineering technologies

我们的目标是制造一种迷你设备，通过包含活细胞来检测化学武器，这些活细胞被设计成在有害的化学战剂存在的情况下变得荧光绿色。该设备将使细胞无限期存活，并能够检测它们产生的荧光绿色的水平，以测量化学武器的浓度。第二次世界大战遗留下来的化学武器在欧洲大陆仍然是一种危险，今天，包括比利时、法国、德国和联合王国在内的几个国家的政府仍然积极参与寻找和销毁旧武器和未爆弹药。在欧洲以外的许多国家，从发达国家(美国)到发展中国家(安哥拉)，通常都保留着以吨位衡量的化学武器能力。因此，越来越需要以足够敏感、有力和可靠的方式检测化学战剂的存在，以保护尽可能多的生命。目前，军方用来检查化学武器存在的大多数设备的工作原理是使用非生物材料，如小型化学品、DNA或抗体，以类似于怀孕测试的方式检测化学武器。这种基于分子的方法的缺点是灵敏度、保质期和灵活性有限，这是因为需要经常将特定化学物质的识别和同一分子内的检测结合起来。活细胞能够以极高的灵敏度探测化合物，最著名的例子是嗅探犬，但据报道，熊可以探测18英里外的食物，即使它们高度敏感的嗅觉也黯然失色。我们寻求通过在我们建议的检测设备中实际驻留活细胞来直接利用细胞的检测能力。我们将首次整合三种不同的技术来建立一种“活的生物传感器”设备：-生物反应器允许细胞以一种精确定义的方式生长和刺激，提供一个封闭的、可控的环境。在制造荷尔蒙或抗体等生物药物时，这种方法被大规模用于将细胞培养到高密度。我们有微型化的生物反应器，可以在台式机或笔记本电脑大小的设备上添加受控环境的细胞生长室。-细胞内的基因模式现在可以重新设计，以形成网络，赋予细胞设计师选择的能力。我们建议设计这样一种基因网络，允许活细胞作为台式机或笔记本电脑大小的设备的组件发挥作用。“合成”基因网络将使细胞能够对化学战剂的存在做出反应，产生一种荧光绿色蛋白，或在未来产生最适合检测设备总体设计的任何其他反应。-微型生物反应器还需要微型管道和电子设备，使其能够保持细胞活着，并可靠地检测荧光绿色蛋白的水平。这需要尖端的微尺度制造技术才能首次制造出如此规模的元件。将这三种方法结合在一个统一的目标--改进世界范围内的化学武器检测--是一种新的方法，我们的努力所产生的技术见解可能会巩固这些技术在医疗诊断和检测食品、药品或服装制造中的污染等领域的许多其他新应用。

We aim to make a mini device that can detect chemical weapons by containing living cells that have been designed to turn fluorescent green in the presence of harmful chemical warfare agents. The device will both keep the cells alive indefinitely and also be able to detect the level of fluorescent green colour they produce in order to measure the concentration of the chemical weapons. Chemical weapons remain a danger in mainland Europe as a legacy of World War II and today the governments of several countries, including Belgium, France, Germany, and the United Kingdom remain actively engaged in locating and destroying old weapons and unexploded ordnance. Outside Europe many countries, ranging from developed (USA) to developing (Angola) typically retain chemical weapons capabilities measured by tonnage. As such there is an increasing need to detect the presence of chemical warfare agents in a manner that is sensitive, robust and reliable enough to safeguard as many lives as possible. Currently most devices used by the military to check for the presence of chemical weapons work by using non-living material such as small chemicals, DNA or antibodies to detect chemical weapons in similar way to how pregnancy tests work. Drawbacks to this 'molecule-based' approach are limited sensitivity, shelf-life and limited flexibility due to the need to often combine recognition of a particular chemical and detection within the same molecule. Living cells can detect compounds with extremely high sensitivity, the most famous example being sniffer dogs, but even their highly-sensitive sense of smell is eclipsed by that of bears who can reportedly detect food 18 miles distant. We seek to directly harness the detection abilities of cells by actually having living cells resident within our proposed detection device. We will, for the first time, integrate three different technologies to build a 'living biosensor' device:- Bioreactors allow cells to be grown and stimulated in a finely defined manner by providing a closed, controlled environment. This is used at large scales to grow cells to high density when making biological medicines such as hormones or antibodies. We have miniaturised bio-reactors down to a 'micro-' scale where controlled-environments cell growth chambers can be added to desktop or laptop-sized devices. - The patterns of genes within cells can now be re-designed to make networks to give cells abilities chosen by the designer. We propose designing such a gene network that allows living cells to function as a component of a desktop or laptop-sized device. The 'synthetic' gene network will enable the cells to respond to the presence of chemical warfare agents by producing a fluorescent green protein or, in future, any other response that best suits the overall design of the detection device. - The miniaturised, 'micro-scale' bioreactor will also require microscale plumbing and electronics to enable it to keep the cells alive and reliably detect levels of the fluorescent green protein. This requires cutting-edge micro-scale fabrication technology to make components at this scale for the first time. Combining these three approaches in a unified goal of improved worldwide chemical weapons detection is a new approach and the technical insights that come from our efforts is likely to underpin many other new applications of these technologies in areas such as medical diagnosis and detection of contamination in food, medicines or clothes manufacture.