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:
  • 金额:
    $ 47.36万
  • 依托单位:
  • 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

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(申请人提供):在这个项目中,MicroFab将设计和制造一个基于喷墨的蛋白质晶体筛选平台原型(CrystalJet(Tm)),重点是难以结晶但对细胞功能至关重要的膜蛋白质。原型平台将进行高通量的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。用于计算和基于结构的药物开发的信息通常是通过对结晶的膜蛋白进行X射线衍射研究而获得的。然而,膜蛋白的结晶往往不能产生衍射性晶体。膜蛋白结晶的困难可以归因于亚稳溶解区狭窄和纯化蛋白的稀少。这些问题的一个解决方案是减少蛋白质消耗,并通过使用较小的筛选体积来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从皮升到微升的小容量的技术,后者是通过吸积实现的。CrystalJet(Tm)筛选平台将在微批和蒸汽扩散模式下运行,以提供实验灵活性。我们还将开发一种一次性喷墨分配器,以解决交叉污染问题。将开发一种DIP-n-Sip模式,用于沉积蛋白质和体积为lt;1000nL的结晶/沉淀剂溶液。笛卡尔流体处理系统将处理更大体积的&gt;1000 NL的流体,如油和油井沉淀剂溶液。该平台将包括用于操纵和密封晶体屏蔽板的板材处理机和板材封口机,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(例如,溶菌酶、葡萄糖异构酶、thaumatin)和膜蛋白质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,可以执行更多的蛋白质结晶学筛选实验,同时与传统筛选系统相比,使用的蛋白质明显更少。最终,这一工具将能够更快、更有效地发现药物靶标,以帮助开发有效的治疗疾病的药物。

项目成果

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Patrick W Cooley其他文献

Patrick W Cooley的其他文献

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{{ truncateString('Patrick W Cooley', 18)}}的其他基金

Inkjet Based System for Lipid Cubic Phase Crystal Screening of GPCR's
用于 GPCR 脂质立方相晶体筛选的喷墨系统
  • 批准号:
    8220753
  • 财政年份:
    2011
  • 资助金额:
    $ 47.36万
  • 项目类别:
Inkjet Based System for Lipid Cubic Phase Crystal Screening of GPCR's
用于 GPCR 脂质立方相晶体筛选的喷墨系统
  • 批准号:
    8056712
  • 财政年份:
    2011
  • 资助金额:
    $ 47.36万
  • 项目类别:
Ink-jet Matrix Coating System for MALDI MS Imaging
用于 MALDI MS 成像的喷墨矩阵涂层系统
  • 批准号:
    7270933
  • 财政年份:
    2007
  • 资助金额:
    $ 47.36万
  • 项目类别:
Ink-Jet System for Protein Crystallization
蛋白质结晶喷墨系统
  • 批准号:
    6933254
  • 财政年份:
    2005
  • 资助金额:
    $ 47.36万
  • 项目类别:
Ink-Jet System for Protein Crystallization
蛋白质结晶喷墨系统
  • 批准号:
    7479601
  • 财政年份:
    2005
  • 资助金额:
    $ 47.36万
  • 项目类别:
OLIGONUCLEOTIDE MICROARRAYS FOR GENE EXPRESSION
用于基因表达的寡核苷酸微阵列
  • 批准号:
    6294235
  • 财政年份:
    2001
  • 资助金额:
    $ 47.36万
  • 项目类别:
LC Microdispenser for High Resolution MALDI-TOF Analysis
用于高分辨率 MALDI-TOF 分析的 LC 微量分配器
  • 批准号:
    6951484
  • 财政年份:
    2000
  • 资助金额:
    $ 47.36万
  • 项目类别:

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