Ink-Jet System for Protein Crystallization

蛋白质结晶喷墨系统

基本信息

批准号：
7479601
负责人：
Patrick W Cooley
金额：
$ 37.62万
依托单位：
MICROFAB TECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-01 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7479601
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）：在该项目中，MicroFab 将设计和制造基于喷墨的蛋白质晶体筛选平台原型（CrystalJet(tm)），重点关注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初始蛋白质结晶筛选，以确定后续放大到衍射质量晶体的参数。基于计算和结构的药物开发信息通常通过结晶膜蛋白的 X 射线衍射研究获得。然而，膜蛋白的结晶通常无法产生衍射质量的晶体。膜蛋白结晶的困难可归因于狭窄的亚稳态溶解区域和纯化蛋白的稀缺。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采样范围。喷墨点胶是一种能够可靠地输送从皮升到微升的小体积的技术，后者是通过累积来实现的。 CrystalJet(tm) 筛选平台将以微批量和蒸汽扩散模式运行，以提供实验灵活性。我们还将开发一次性喷墨分配器来解决交叉污染问题。将开发一种浸入-n-sip 模式，用于沉积蛋白质并匹配体积<1000 nL 的结晶/沉淀溶液。笛卡尔流体处理系统将操纵 >1000 nL 的较大流体体积，例如油和井沉淀剂溶液。该平台将包括用于操纵和密封晶体筛选板的板处理机和板密封器、用于保持温度、湿度和清洁空气的环境控制，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）和膜蛋白进行一系列筛选实验，以验证 CrystalJet(tm) 平台的运行。 CrystalJet 平台是一种自动化工具，可进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，使用的蛋白质显着减少。最终，该工具将实现更快、更有效的药物靶点发现，以帮助开发治疗疾病的有效药物。