Zhu Ying, Zhu Li-Na, Guo Rui, Cui Heng-Jun, Ye Sheng, Fang Qun
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China.
Life Sciences Institute, Zhejiang University, Hangzhou, 310058, China.
Sci Rep. 2014 May 23;4:5046. doi: 10.1038/srep05046.
Large-scale screening of hundreds or even thousands of crystallization conditions while with low sample consumption is in urgent need, in current structural biology research. Here we describe a fully-automated droplet robot for nanoliter-scale crystallization screening that combines the advantages of both automated robotics technique for protein crystallization screening and the droplet-based microfluidic technique. A semi-contact dispensing method was developed to achieve flexible, programmable and reliable liquid-handling operations for nanoliter-scale protein crystallization experiments. We applied the droplet robot in large-scale screening of crystallization conditions of five soluble proteins and one membrane protein with 35-96 different crystallization conditions, study of volume effects on protein crystallization, and determination of phase diagrams of two proteins. The volume for each droplet reactor is only ca. 4-8 nL. The protein consumption significantly reduces 50-500 fold compared with current crystallization stations.
在当前的结构生物学研究中，迫切需要在低样品消耗的情况下对数百甚至数千种结晶条件进行大规模筛选。在此，我们描述了一种用于纳升规模结晶筛选的全自动液滴机器人，它结合了用于蛋白质结晶筛选的自动化机器人技术和基于液滴的微流控技术的优点。开发了一种半接触式分配方法，以实现用于纳升规模蛋白质结晶实验的灵活、可编程且可靠的液体处理操作。我们将该液滴机器人应用于对五种可溶性蛋白质和一种膜蛋白的35 - 96种不同结晶条件的结晶条件大规模筛选、蛋白质结晶体积效应的研究以及两种蛋白质相图的测定。每个液滴反应器的体积仅约4 - 8纳升。与当前的结晶工作站相比，蛋白质消耗量显著降低了50 - 500倍。
