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

该项目将开发一个基于单细胞微流控递送的超高通量筛选平台1，并结合质谱分析2。您将应用此快速筛选合成生物学文库，以识别和定量小分子靶标。液滴将被引导至给定质谱仪的电喷雾电离源，用于随后的快速质量分析，从而能够进行鉴定，并确定给定文库中给定细胞类型的“成功”。最终，该项目的目标将是使用picodroplet驱动的质谱分析来靶向命中化合物，并开发流体控制，以从中学习，然后触发相同的picodroplet分选到分配器中进行后续下游处理。在曼彻斯特生物技术研究所的SYNBIOCHEM中心工作，您将开发和优化这种新型微流体平台与质谱仪的接口。picodroplet装置已经被证明能够在5.4小时内筛选200，000个picodroplet，这可能允许在一天内分析2-4个完整的文库。我们将集中在图书馆设计产生萜烯和黄酮类化合物的初步分析，虽然我们预计，这项技术将有广泛的使用在一些系统。利用我们在蛋白质质谱3方面的专业知识，我们还将开发必要的电离和液滴分配方案，以直接从细胞中分析经工程改造以制造小分子靶标的蛋白质。通过这种方式，该平台将用于优化库设计和构建，并形成我们合成生物学中心的一个组成部分。

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.