Characterisation of cellular assemblies in microfluidic systems (synthetic biology to obtain novel antibiotics and optimized production systems)

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The proposed research project forms an integral part of the larger international proposal SYNMOD (Full name: Synthetic biology to obtain novel antibiotics and optimised production systems), which is a collaborative project with five partners in Europe and one in the UK. SYNMOD seeks to integrate the basic science of Synthetic Biology with the necessary bioprocess engineering in order to achieve new and improved antibiotics to overcome the growing problem of antimicrobial resistance. In order to realise this potential, it is necessary to undertake research to develop protocols and technologies that allow to quickly and easily characterise biological parts and cells, and to understand their behaviour in different growth conditions. Developing such technologies and protocols is the goal of this proposed research project. Currently, some tools, such as shaker flasks, are easy to set up for an experiment, but provide relatively little information per experiment. Other tools, such as bench-scale bioreactors with multiple probes, are costly and labour-intensive to set up for an experiment. It is therefore difficult to perform the large number of experiments required to fully understand the behaviour of biological parts and cells. The proposed research addresses this by using micro-fabrication methods to create chambers with gas permeable membranes to provide the oxygen cells need. Through such 'microfluidic' reactor systems for the cultivation of cells have been fabricated, they have mainly been designed to operate in 'batch' mode. This is a reactor operation mode where the culture conditions vary considerably during an experiment. Accordingly, the properties of the cells, such as size, composition, and functional characteristics vary considerably, too. In contrast, in a 'chemostat' operation mode, the cells are kept in the same growth condition, i.e. the cell biomass, the nutrient and the product concentrations in the culture remain constant. This is achieved by a controlled and continuous supply of nutrient combined with a controlled and continuous removal of product and of 'excess' cells. Therefore, in a 'chemostat' operation mode, it becomes possible to correlate a cellular behaviour with a particular growth condition. In this research, we propose to build 'microfluidic' reactor systems that operate in 'chemostat' mode. Once this is solved, it will then become possible to use the general advantages of 'microfluidic' systems, such as parallelisation and automation, to create a cost-effective tool with which cell behaviour can be rapidly investigated and characterised.