A single microfluidic chip will sort, fuse, and analyse thousands of water droplets per second, each one a tiny chemical reactor searching for new enzymes. Standard methods for discovering novel biological catalysts are slow and labour-intensive, often requiring separate experiments for each candidate. This project builds a modular lab-on-a-chip that automates the entire process—loading droplets with reaction components, fusing them to add new compounds, incubating them, and then screening for products using either spectroscopy or mass spectrometry. The mass spectrometry interface is particularly challenging because no one has reliably coupled it to fast-moving microdroplets before. If the device works, it will let researchers run directed evolution experiments on enzymes far more efficiently than existing approaches allow. That could accelerate the discovery of enzymes with new or improved functions—catalysts that might one day drive greener chemical manufacturing, break down plastic waste, or synthesise pharmaceuticals with fewer byproducts. Because the platform is modular, it can be customised for problems across biology, chemistry, and materials science. The project is fundamentally about building a tool, not about any single application, but the tool itself could make previously impractical experiments routine.
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We plan to generate a novel platform technology for experimental science, and demonstrate its utility by applying it to the identification of novel biological catalysts for chemical reactions. The key features of the technology will be its speed, scale and general utility. Individual reactions will take place inside microdroplets of water carried in a fluorocarbon continuous phase within microfluidic channels. These microreactors will be load up with the components for a reaction. Additional compounds can then be added by fusing a droplet with a second droplet. Once a reaction has occurred in a droplet the presence of product will be determined either spectroscopically or by mass spectrometry. Mass spectrometry has the advantage that it can provide high resolution structural information. However interfacing it with microdroplets is an unexplored and very challenging part of the proposal. In this way new catalysts will be identified which can then be improved by taking them through several rounds of selection.A modular device will be assembled that load, fuse, incubate sort and split droplets and present then to either a spectroscopic or mass spectrometric screen. The fabrication of this device, and especially the integration of the various components is a significant challenge, and will be the main focus of the first part of the project.We plan to use this modular device for the discovery of novel biological catalysts. These will either be enzymes that incorporate new functionality, or enzymes from unusual organisms. This is an example of an important project that will become more accessible using the system we develop than using than with existing approaches. We expect to get an efficiency enhancement of over 104. The project is multidisciplinary and at the forefront of several areas: the use of microdroplets, the use of microfluidics for lab-on-a-chip applications, the use of mass spectrometry, and the directed evolution of enzymes. It will be possible to customise the devices we generate to tackle a broad range of problems in biological, chemical and materials science.
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