Development of a Ultra High Throughput Screening Platform for Synthetic Biology - From Cells to Validated Target

该项目将开发一种基于单细胞微流控递送的超高通量筛选平台1与质谱分析2。您将把它应用于快速筛选合成生物学文库，以识别和量化小分子靶标。液滴将被定向到给定质谱计的电喷雾电离源，用于随后的快速质量分析，从而能够进行鉴定，并确定给定的细胞类型在给定的库中的“成功”。最终，该项目的目标是使用微滴驱动的质谱分析来瞄准命中化合物，并开发流体控制以学习这一点，然后触发将相同的微滴分选到分配器进行后续处理。在我们SYNBIOCHEM中心的曼彻斯特生物技术研究所工作，您将开发和优化这一新型微流控平台与质谱仪的接口。微滴装置已经被证明能够在5.4小时内筛选20万微滴，这可能允许在一天内分析2-4个完整的文库。该筛选平台将被应用于SYNBIOCHEM文库的分析。我们将把初步分析的重点放在为产生萜类和类黄酮而设计的文库上，尽管我们预计这项技术将在许多系统中广泛使用。利用我们在蛋白质质谱学3方面的专业知识，我们还将开发必要的电离和液滴消散方案，以直接从细胞中分析为制造小分子靶标而设计的蛋白质。通过这种方式，这个平台将被用来优化图书馆的设计和建设，并形成我们合成生物学中心不可或缺的一部分。

This project will develop an ultra-high throughput screening platform based on microfluidic delivery of single cells1 coupled to mass spectrometry analysis2. You will apply this to rapidly screen synthetic biology libraries to identify and quantify small molecule targets. Droplets will be directed to the electrospray ionisation source of a given mass spectrometer, for subsequent rapid mass analysis enabling identification, and to determine the 'success' of a given cell type in a given library. Ultimately, this project will aim to use the picodroplet driven mass spectrometry analysis to target hit compounds and develop the fluidics control to learn from this to then trigger the sorting of identical picodroplets to a dispenser for subsequent downstream processing.Working in the Manchester Institute of Biotechnology in our SYNBIOCHEM centre you will develop and optimise the interface of this novel microfluidic platform to a mass spectrometer. The picodroplet device has already been shown to be capable of screening 200,000 picodroplets in 5.4 hours, which potentially would allow 2-4 entire libraries to be assayed in a day.This screening platform will be applied to the analysis of SYNBIOCHEM libraries. We will focus initial analysis on libraries designed to generate terpenes and flavonoids, although we anticipate that this technology will have wide use on a number of systems. Using our expertise in protein mass spectrometry3 we will also develop the necessary ionisation and droplet dispension protocols to also analyse directly from cells the proteins that have been engineered to make the small molecule targets. In this way this platform will be used to optimise library design and build and form an integral part of our synthetic biology centre.