GOALI: Biocatalysis Development using High-throughput Droplet Microfluidics and Mass Spectrometry

1604087罗伯特·肯尼迪 该项目的目标是开发一种新的方法，以促进更快，更环保，更便宜的合成药物的方法的发展。 使用酶的挑战在于天然存在的酶通常不催化药物开发项目所需的确切反应。该项目的目标是开发一种高通量，高度小型化的工程酶方法来执行所需的反应。预计该技术可用于在几天内开发此类生物催化剂，用于任何所需的化学转化。这一能力可以以时间和成本效益高的方式为任何反应提供量身定制的催化剂，从而大大促进药物开发项目。由于酶促转化可能提供高收率、高选择性和高对映体纯度、低成本和绿色化学，因此用于制药过程化学和制造的酶工程已成为一种革命性技术。该团队建议开发一种新的筛选系统，以减少材料使用，消除特殊试剂，提高酶工程和筛选的整体通量。三个关键模块包括（i）无细胞蛋白质合成（CFPS），（ii）液滴微流体和（iii）质谱（MS）。CFPS提供了潜在的速度优势，因为它绕过了细胞生长和蛋白质收获步骤。它可以在更短的周期内完成，生产更纯的蛋白质，同时允许主动监测，直接操作和快速采样。液滴微流体技术提供了一个平台，用于在纳升或更小的体积内进行反应和测定，这也适合于自动化。MS允许以最小的延迟快速开发用于各种潜在底物的无标记测定的可能性。所提出的方法是变革性的，因为它将允许几乎任何酶促转化在10倍的时间内通过消除开发标记，偶联反应或在过程中使用细胞的需要而进行工程化。该技术也将用于化学过程开发的其他应用。该奖项由CBET部门的生物技术和生物化学工程计划共同资助，由工业创新和伙伴关系部门的GOALI计划共同资助。

1604087Kennedy, Robert T. The goal of this project is to develop a new approach to facilitate the development of faster, greener, and cheaper ways of synthesizing drugs. The challenge in using enzymes is that natural occurring enzymes do not often catalyze the exact reaction that is needed for a pharmaceutical development project. The goal in this project is to develop a high-throughput, highly miniaturized method of engineering enzymes to perform desired reactions. It is anticipated that this technology can be used to develop such biocatalysts in a few days for any desired chemical transformation. This capability could greatly facilitate drug development projects by making available tailor-made catalysts for any reaction in a time and cost-effective manner. Enzyme engineering for pharmaceutical process chemistry and manufacturing has become a revolutionary technology as enzymatic transformations potentially offer high yield, selectivity and enantiomeric purity with low cost and green chemistry. The team proposes development of a novel screening system to reduce the material use, eliminate special reagents and improve the overall throughput in enzyme engineering and screening. The three key modules include (i) cell-free protein synthesis (CFPS), (ii) droplet microfluidics and (iii) mass spectrometry (MS). CFPS offers the potential speed advantage as it bypasses the cell growth and protein harvest steps. It can be completed with shorter cycle time, produces purer proteins, and at the same time allows active monitoring, direct manipulation, and rapid sampling. Droplet microfluidics offers a platform for performing reactions and assays in nanoliter or less volume which is also adaptive to be automated. MS allows the possibility for fast development of label-free assays for a wide variety of potential substrates with minimal delay. The proposed approach is transformative because it will allow virtually any enzymatic transformation to be engineered in 10-fold less time by eliminating the need to develop labels, coupled reactions, or using cells in the process. The proposed technology will be useful also for other applications in chemical process development.This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the GOALI Program of the Division of Industrial Innovation and Partnerships.