The Crystal Optimizer: Kinetic Control of Protein Crystallization

晶体优化器：蛋白质结晶的动力学控制

• 批准号: 7404268
• 负责人: DARREN R LINK
• 金额: $ 10万
• 依托单位: RAINDANCE TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-15 至 2009-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7404268
• 关键词: Blood capillaries; Budgets; Chemicals; Condition; Crystallization; Device Designs; Devices; Drops; Engineering; Equilibrium; Funding; Goals; Growth; Image; Individual; Kinetics; Lead; Measures; Methods; Microfluidic Microchips; Modeling; Pathway interactions; Phase; Plumbing; Process; Production; Protein Structure Initiative; Proteins; Pump; Range; Resources; Sampling; Screening procedure; Siloxanes; Solutions; Staging; Standards of Weights and Measures; Structure; Students; Surface; Syringes; Technology; Temperature; Testing; Time; Wages; Work; capillary; concept; design; improved; innovation; instrument; instrumentation; manufacturing process; protein structure; prototype; solute; structural biology

DESCRIPTION (provided by applicant): Protein crystallization remains a bottleneck in the goal to determine the protein's structure. A microfluidic device denoted the Crystal Optimizer (XOpt) has been developed to optimize the kinetic pathway for the nucleation and growth of protein crystals once the lead conditions have been identified by other screening methods. The device drives each of hundreds of drops of the candidate protein solution to different degrees of supersaturation for variable durations and then reduces the supersaturation to a low value. The concept is to find the two-step kinetic pathway that firstly quenches the drop for the right amount of time to nucleate just a few crystals and secondly reduces the supersaturation to a level that suppresses nucleation of additional crystals, but is still supersaturated enough for the crystals to grow slowly. Besides controlling concentration, the XOpt measures the solute and protein concentrations as a function of time so quantitative information on the actual kinetic pathway traversed by the samples can be obtained and exploited to design improved kinetic pathways in subsequent rounds of optimization. Addition of a simple temperature stage that generates a thermal gradient across the chip is used to find the optimal temperature of the optimal kinetic path. Two key points make the Crystal Optimizer innovative and useful. First, protein crystallization is an activated, non-equilibrium process and the XOpt is the sole microfluidic device designed to systematically optimize this physical feature in a controlled and reversible manner. Second, the XOpt is simple to use and inexpensive to manufacture.

描述（由申请人提供）：蛋白质结晶仍然是确定蛋白质结构目标的瓶颈。已经开发了一种称为晶体优化器（XOpt）的微流体装置，以在通过其他筛选方法确定了先导条件后优化蛋白质晶体的成核和生长的动力学途径。该装置驱动数百滴候选蛋白质溶液中的每一滴在可变的持续时间内达到不同程度的过饱和，然后将过饱和度降低到低值。这个概念是找到两步动力学途径，首先将液滴淬灭适量的时间以使仅几个晶体成核，其次将过饱和度降低到抑制额外晶体成核的水平，但仍足够过饱和以使晶体缓慢生长。除了控制浓度外，XOpt还测量溶质和蛋白质浓度随时间的变化，因此可以获得样品实际动力学途径的定量信息，并在后续优化中用于设计改进的动力学途径。添加一个简单的温度阶段，产生一个热梯度的芯片是用来找到最佳的动力学路径的最佳温度。 两个关键点使Crystal Optimizer具有创新性和实用性。首先，蛋白质结晶是一个激活的非平衡过程，XOpt是唯一的微流体设备，旨在以可控和可逆的方式系统地优化这一物理特征。其次，XOpt使用简单，制造成本低廉。