Ink-Jet System for Protein Crystallization

蛋白质结晶喷墨系统

• 批准号: 6933254
• 负责人: Patrick W Cooley
• 金额: $ 10.85万
• 依托单位: MICROFAB TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6933254
• 关键词: X ray crystallography; bioimaging /biomedical imaging; detergents; high throughput technology; membrane proteins; nanotechnology; protein structure; structural biology; technology /technique development

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.

描述（由申请人提供）：在该项目中，MicroFab将开发一种基于喷墨的高通量系统，以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屏幕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的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，完整膜蛋白的结晶通常不能产生可用于衍射分析的晶体。膜蛋白（MP）结晶困难可归因于其非常窄的亚稳溶解度区域和材料稀缺。这些问题的共识解决方案是通过使用较小的筛选体积来减少蛋白质消耗并缩小晶体生长条件的取样范围。喷墨点胶是一种能够可靠地提供从皮升到微升的小体积的技术，后者通过增加。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溶液所需的操作参数。MicroFab还将确定在油下聚结和混合这些溶液的最佳工艺。最后，MicroFab将使用喷墨点胶进行膜蛋白的初始结晶条件筛选。我们期望这些实验提供关键信息，最终使高通量皮升MP晶体生长测定成为主流技术。