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

采用微型恒化器和细胞工程技术的自我再生悬浮相全细胞生物传感器系统

• 批准号: BB/J020605/1
• 负责人: Nicolas Szita
• 金额: $ 9.73万
• 依托单位: University 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=BB%2FJ020605%2F1
• 关键词: Self; regenerating; suspended; phase; whole

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.

我们的目标是制造一种微型装置，通过含有活细胞来检测化学武器，这些活细胞被设计成在有害化学战剂存在的情况下变成荧光绿色。该装置将使细胞无限期地存活，并能够检测它们产生的荧光绿色水平，以测量化学武器的浓度。化学武器作为第二次世界大战的遗产在欧洲大陆仍然是一种危险，今天包括比利时、法国、德国和英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。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，越来越需要以足够灵敏、稳健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，军方用于检查化学武器存在的大多数设备都是通过使用非生物材料，如小型化学品，DNA或抗体来检测化学武器，其工作方式与怀孕测试的工作方式类似。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，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进行联合收割机。活细胞可以以极高的灵敏度检测化合物，最著名的例子是嗅探犬，但即使是它们高度敏感的嗅觉也被熊黯然失色，据报道熊可以检测到18英里外的食物。我们试图直接利用细胞的检测能力，实际上有活细胞居住在我们提出的检测设备。我们将首次整合三种不同的技术来构建一个“活体生物传感器”设备：-生物反应器通过提供一个封闭的、受控的环境，允许细胞以一种精细定义的方式生长和刺激。这是用于大规模生长细胞高密度时，制造生物药物，如激素或抗体。我们已经将生物反应器简化到“微”尺度，其中受控环境细胞生长室可以添加到桌面或笔记本电脑大小的设备中。- 细胞内的基因模式现在可以被重新设计，以形成网络，从而赋予细胞由设计者选择的能力。我们建议设计这样一个基因网络，使活细胞作为桌面或笔记本电脑大小的设备的组成部分。“合成”基因网络将使细胞能够通过产生荧光绿色蛋白质来对化学战剂的存在做出反应，或者在未来，任何其他最适合检测设备整体设计的反应。- 这种“微型”生物反应器还需要微型管道和电子设备，使其能够保持细胞存活，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来首次制造这种规模的组件。将这三种方法结合在一个改进全世界化学武器检测的统一目标中是一种新的方法，我们的努力所产生的技术见解可能会支持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。

项目成果

Electromagnetic stirring in a microbioreactor with non-conventional chamber morphology and implementation of multiplexed mixing.

• DOI: 10.1002/jctb.4762
• 发表时间: 2015-10
• 期刊: Journal of chemical technology and biotechnology (Oxford, Oxfordshire : 1986)
• 作者: Tan CK;Davies MJ;McCluskey DK;Munro IR;Nweke MC;Tracey MC;Szita N
• 通讯作者: Szita N

Oxygen transfer characteristics of miniaturized bioreactor systems.

微型生物反应器系统的氧转移特性。

• DOI: 10.1002/bit.24824
• 发表时间: 2013-04
• 期刊: BIOTECHNOLOGY AND BIOENGINEERING
• 影响因子: 3.8
• 作者: Kirk, Timothy V.;Szita, Nicolas
• 通讯作者: Szita, Nicolas

Development of a microbioreactor 'cassette' for the cultivation of microorganisms in batch and chemostat mode

开发用于以批量和恒化器模式培养微生物的微生物反应器“盒”

• 发表时间: 2013
• 期刊: Chimica Oggi - Chemistry Today
• 作者: Davies, MJ