Ink-Jet System for Protein Crystallization

蛋白质结晶喷墨系统

基本信息

批准号：
7324044
负责人：
Patrick W Cooley
金额：
$ 47.36万
依托单位：
MICROFAB TECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-10-01 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7324044
关键词：
Address Affinity Area Aspirate substance Automation Back Caliber Cell physiology Classification Compatible Complex Computer software Condition Consensus Consumption Crystallization Crystallography Cultured Cells Defect Deposition Detergents Development Devices Diffuse Diffusion Dimensions Disease Drops Drug Delivery Systems Drug Design Endopeptidases Equilibrium Evaluation Excision Exhibits Generations Growth Hour Humidity Image Individual Informatics Ink Integral Membrane Protein Investigation Kinetics Liquid substance Membrane Membrane Proteins Methods Microscopy Molecular Muramidase Names Numbers Oils Operative Surgical Procedures Particulate Peptide Hydrolases Performance Pharmaceutical Preparations Phase Phospholipids Plant Resins Pliability Polytetrafluoroethylene Printing Process Production Productivity Protein Denaturation Protein Overexpression Proteins Protocols documentation Range Rate Reagent Recombinant Proteins Research Research Personnel Robot Running Sampling Screening procedure Series Solubility Solutions Solvents Staging Standards of Weights and Measures Structure Synchrotrons System Technology Temperature Test Result Testing Thinking Tube Validation Vision Week X-Ray Crystallography air cleaner base commercialization cost day design design and construction dodecyl maltoside dodecyldimethylamine oxide drug development experience fluid flow glucose isomerase improved interest macromolecule milligram nanolitre nanoscale numb protein octylglucopyranoside pressure prevent protein structure prototype research study scale up seal size small molecule structural biology success tool trend vapor

项目摘要

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.

描述（由申请人提供）：在此项目中，Microfab将设计和制造基于原型喷墨的蛋白质晶体筛选平台（CrystalJet（TM）），重点是难以结晶的膜蛋白，但对细胞功能至关重要。原型平台将执行高通量初始蛋白质结晶筛选，以识别参数，以便随后扩展到衍射质量晶体。通常通过结晶膜蛋白的X射线衍射研究获得基于计算和结构的药物发育的信息。但是，膜蛋白的结晶通常无法产生衍射质量晶体。结晶膜蛋白的困难可能归因于狭窄的亚稳态溶解度区域和纯化蛋白的稀缺性。解决这些问题的一种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采样。墨水分配是一种能够可靠地提供少量量的技术，从Picoliter到Microliter，后者通过增生。 CrystalJet（TM）筛选平台将以微匹配和蒸气扩散模式运行，以提供实验灵活性。我们还将开发一次一次性喷墨分配器来解决交叉污染问题。将开发出一种浸入N- SIP模式，用于在小于1000 nl的体积下蛋白质的沉积和结晶/沉淀物溶液的匹配体积。笛卡尔流体处理系统将操纵> 1000 nl的较大流体体积，例如油和淀粉溶液。该平台将包括一个平板处理程序和板密封剂，以操纵和密封晶体屏幕板，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，thaumatin）和膜蛋白来验证晶体平台的运行（TM）平台的运行，将进行一系列筛选实验。 CrystalJet平台是一种自动化工具，可以执行更多的蛋白质晶体学筛选实验，同时与常规筛选系统相比，使用明显较少的蛋白质。最终，该工具将使发现更快，更有效的药物靶标的发现，以帮助开发有效的药物治疗疾病。