Hansen Carl L, Classen Scott, Berger James M, Quake Stephen R
Department of Applied Physics, California Institute of Technology, 1200 East California Boulevard, Pasadena, California 91125, USA.
J Am Chem Soc. 2006 Mar 15;128(10):3142-3. doi: 10.1021/ja0576637.
The unprecedented economies of scale and unique mass transport properties of microfluidic devices made them viable nano-volume protein crystallization screening platforms. However, realizing the full potential of microfluidic crystallization requires complementary technologies for crystal optimization and harvesting. In this paper, we report a microfluidic device which provides a link between chip-based nanoliter volume crystallization screening and structure analysis through "kinetic optimization" of crystallization reactions and in situ structure determination. Kinetic optimization through systematic variation of reactor geometry and actuation of micromechanical valves is used to screen a large ensemble of kinetic trajectories that are not practical with conventional techniques. Using this device, we demonstrate control over crystal quality, reliable scale-up from nanoliter volume reactions, facile harvesting and cryoprotectant screening, and protein structure determination at atomic resolution from data collected in-chip.
微流控装置前所未有的规模经济和独特的质量传输特性使其成为可行的纳升级蛋白质结晶筛选平台。然而，要充分发挥微流控结晶的潜力，还需要用于晶体优化和收获的互补技术。在本文中，我们报告了一种微流控装置，该装置通过结晶反应的“动力学优化”和原位结构测定，在基于芯片的纳升体积结晶筛选和结构分析之间建立了联系。通过系统改变反应器几何形状和驱动微机械阀进行动力学优化，用于筛选大量传统技术无法实现的动力学轨迹。使用该装置，我们展示了对晶体质量的控制、从纳升体积反应可靠放大、轻松收获和冷冻保护剂筛选，以及从芯片内收集的数据以原子分辨率确定蛋白质结构。
