Ink-Jet System for Protein Crystallization

描述（由申请人提供）：在该项目中，MicroFab将设计和制造一个基于喷墨的蛋白质晶体筛选平台（CrystalJet（TM）），重点是难以结晶但对细胞功能至关重要的膜蛋白。该原型平台将进行高通量的初始蛋白质结晶筛选，以确定随后放大到衍射质量晶体的参数。用于基于计算和结构的药物开发的信息通常通过结晶膜蛋白的X射线衍射研究获得。然而，膜蛋白的结晶通常不能产生衍射质量的晶体。结晶膜蛋白的困难可归因于狭窄的亚稳溶解度区域和缺乏纯化的蛋白质。这些问题的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范围内的小体积的技术，后者通过增加。CrystalJet（TM）筛选平台将以微批次和蒸汽扩散模式运行，以提供实验灵活性。我们亦将开发一次性喷墨分配器，以解决交叉污染问题。将开发dip-n- sip模式，用于蛋白质沉积和体积为1000 nL的结晶/沉淀剂溶液<1000 nL. A Cartesian fluid handling system will manipulate larger fluid volumes of >（如油和孔沉淀剂溶液）的匹配体积。该平台将包括一个板处理器和板处理器，用于操作和密封晶体筛选板，环境控制，以保持温度，湿度和清洁空气，以及晶体观察和图像捕获系统。将使用蛋白质标准品（例如，溶菌酶、葡萄糖异构酶、奇异果甜蛋白）和膜蛋白以验证CrystalJet（TM）平台的操作。CrystalJet平台是一种自动化工具，可执行更多数量的蛋白质晶体学筛选实验，同时与传统筛选系统相比，使用的蛋白质显著减少。最终，该工具将能够更快，更有效地发现药物靶点，以帮助开发治疗疾病的有效药物。

DESCRIPTION (provided by applicant): In this project, MicroFab will design and fabricate a prototype ink-jet based protein crystal screening platform (CrystalJet(tm)) with emphasis on membrane proteins that are difficult to crystallize, but critical to cellular function. The prototype platform will perform high-throughput initial protein crystallization screening to identify parameters for subsequent scale-up to diffraction quality crystals. Information for computational and structure based drug development is typically obtained through x-ray diffraction studies of crystallized membrane proteins. However, the crystallization of membrane proteins often fails to produce diffraction quality crystals. Difficulties in crystallizing membrane proteins can be attributed to narrow metastable solubility regions and scarcity of purified protein. A 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 latter by accretion. The CrystalJet(tm) screening platform will operate in microbatch and vapor diffusion modes to offer experimental flexibility. We will also develop a disposable ink-jet dispenser to address cross contamination issues. A dip-n- sip mode will be developed for the deposition of protein and matching volumes of crystallization/precipitant solution at volumes of <1000 nL. A Cartesian fluid handling system will manipulate larger fluid volumes of >1000 nL, such as oil and well precipitant solutions. The platform will include a plate handler and plate sealer to manipulate and seal crystal-screening plates, environmental control to maintain temperature, humidity and clean air, and a crystal observation and image capture system. A series of screening experiments will be performed using protein standards (e.g., lysozyme, glucose isomerase, thaumatin) and membrane proteins to validate the operation of the CrystalJet(tm) platform. The CrystalJet platform is an automated tool to perform a greater number of protein crystallography screening experiments, while using significantly less protein in comparison to conventional screening systems. Ultimately, this tool will enable faster and more efficient drug target discovery to aid in the development of effective drugs to treat disease.