Automated microfluidic sample-preparation platform for high-throughput structural investigation of proteins by small-angle X-ray scattering
Automated microfluidic sample-preparation platform for high-throughput structural investigation of proteins by small-angle X-ray scattering
复制标题
DOI：
10.1107/s0021889811030068
复制
发表时间：
2011-10-01
影响因子：
6.1
通讯作者：
Kutter, Jorg P.
中科院分区：
文献类型：
--
作者：
Lafleur, Josiane P.;Snakenborg, Detlef;Kutter, Jorg P.
提出了一种新的微流控样品制备系统，用于在同步加速器上用小角X射线散射（SAXS）研究蛋白质的结构。该系统包括硬件和软件功能，用于精确的流体控制、扩散混合样品、自动X射线曝光控制、紫外吸收测量和自动数据分析。仅需15 μ l样品即可完成一个完整的分析周期，包括样品混合、SAXS测量、连续UV吸光度测量以及用缓冲液清洁通道和X射线池。完整的分析周期可以在不到3分钟内进行。牛血清白蛋白被用作模型蛋白质，以表征系统的混合效率和样品消耗。衔接蛋白（p120-RasGAP）的N2片段被用于演示该设备如何通过使用高通量技术筛选广泛的条件来调查蛋白质的结构空间。
A new microfluidic sample-preparation system is presented for the structural investigation of proteins using small-angle X-ray scattering (SAXS) at synchrotrons. The system includes hardware and software features for precise fluidic control, sample mixing by diffusion, automated X-ray exposure control, UV absorbance measurements and automated data analysis. As little as 15 mu l of sample is required to perform a complete analysis cycle, including sample mixing, SAXS measurement, continuous UV absorbance measurements, and cleaning of the channels and X-ray cell with buffer. The complete analysis cycle can be performed in less than 3 min. Bovine serum albumin was used as a model protein to characterize the mixing efficiency and sample consumption of the system. The N2 fragment of an adaptor protein (p120-RasGAP) was used to demonstrate how the device can be used to survey the structural space of a protein by screening a wide set of conditions using high-throughput techniques.