Ink-Jet System for Protein Crystallization

描述（由申请人提供）：在该项目中，MicroFab将开发一种高通量喷墨系统，用于筛选膜蛋白结晶的初始迹象，同时最大限度地保护有限的蛋白质样品。然后，研究人员可以重复从初始筛选确定的成功结晶实验，以生长更大的有序晶体用于衍射实验。通过x射线晶体学获得的膜蛋白结构可以为基于计算和结构的药物开发提供信息。不幸的是，整体膜蛋白的结晶常常不能产生用于衍射分析的晶体。膜蛋白（MP）的结晶困难可归因于其亚稳溶解度非常窄和材料的稀缺性。解决这些问题的共识是通过使用较小的筛选体积来减少蛋白质消耗和缩小晶体生长条件的采样范围。喷墨点胶是一种能够可靠地提供小体积的技术，从皮升到微升，后者通过吸积。在第一阶段，MicroFab将确定使用喷墨设备分配蛋白质和洗涤剂筛选解决方案所需的操作参数。MicroFab还将确定这些溶液在油下凝聚和混合的最佳工艺。最后，MicroFab将使用喷墨点胶对膜蛋白进行初始结晶条件筛选。我们希望这些实验能够提供关键信息，最终使高通量皮升MP晶体生长分析成为主流技术。

DESCRIPTION (provided by applicant): In this project, MicroFab will develop a high throughput ink-jet based system to screen for the initial signs of crystallization of membrane proteins, while maximizing the conservation of limited protein samples. Investigators can then repeat successful crystallization experiments determined from the initial screens to grow larger well-ordered crystals for diffraction experiments. The structure of membrane proteins obtained by x-ray crystallography can provide information for computational and structure based drug development. Unfortunately, the crystallization of integral membrane proteins often fails to yield crystals useful for diffraction analysis. Membrane protein (MP) crystallization difficulties can be attributed to their very narrow metastable solubility regions and scarcity of material. The consensus solution to these problems is to decrease protein consumption and narrow the sampling of crystal growth conditions by using smaller screening volumes. Ink-jet dispensing is a technology capable of reliably delivering small volumes ranging from picoliter to microliter, the later through accretion. In Phase I MicroFab will determine the operational parameters required to dispense protein and detergent screening solutions using ink-jet devices. MicroFab will also determine the optimum process for coalescing and mixing of these solutions under oil. Finally, MicroFab will conduct initial crystallization condition screens of membrane proteins using ink-jet dispensing. We expect these experiments to provide critical information that will ultimately enable high throughput picoliter MP crystal growth assays as a mainstream technique.