Biomolecule Crystallization Microarray

生物分子结晶微阵列

• 批准号: 6736801
• 负责人: PAUL W TODD
• 金额: $ 79.26万
• 依托单位: TECHSHOT, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6736801
• 关键词: X ray crystallography; biomedical equipment development; bioreactors; crystallization; fluid flow; lysozyme; macromolecule; microarray technology; monitoring device; structural biology

DESCRIPTION (provided by applicant): Several aspects of health and medicinal research depend on the knowledge of macromolecular structures, the determination of which depends on a supply of high-quality crystals. Conditions for growing high-quality crystals are usually determined by expensive trial-and-error research. The proposed innovation is a lithography-fabricated system for automated, kinetically controlled growth of biomolecule crystals for crystallography. It is also useful in crystallization process development applications. An array of crystallization chambers, each with a volume of a few microlitres is served by an array of pumps whose feed rates are controlled in the sub-nL/s range by a time-of-flight flow meter. The pumps, one each for buffer, biomolecule solution and precipitant solution, are commanded by control algorithms that use image data. The individual concepts were demonstrated in Phase I research, namely, the ability to deliver a few microlitres of solution to a microreactor chamber over several hours and to grow crystals in such a chamber, the ability to electronically measure sub nL/s flow rates with a meter than has no moving parts, and the identification of a very inexpensive digital imaging and analysis method. The Phase II research specific aims are to (1) integrate these novel components into a single automated crystallization system to be tested for the coordination of all functions through a single control computer; (2) on the basis of (1) design and fabricate by lithography an integrated 4-chamber array including optics, microfluidics and electronics; and (3) test this array against real crystal-growth problems and refine image-based control algorithms. This technology uses automated kinetic control of the liquid-liquid crystallization method as a means of canvassing a continuum of chemical conditions rather than using massively paralleled discreet screening to enhance throughput in this rather expensive process. The resulting product, a crystallizer microarray ("CRYMA"), will be marketed to the world's large pharmaceutical firms, university laboratories with structural biology units and governmental space research agencies - a market with which SHOT, Inc. is familiar. Its implementation.

描述（由申请人提供）：健康和医学研究的几个方面取决于大分子结构的知识，其确定取决于高质量晶体的供应。生长高质量晶体的条件通常是通过昂贵的试错研究来确定的。提出的创新是一种光刻制造的系统，用于自动化，动力学控制的生物分子晶体生长的晶体学。它在结晶过程开发应用中也很有用。一系列结晶室，每个室的体积为几微升，由一系列泵提供服务，泵的进料速率由飞行时间流量计控制在低于nL/s的范围内。这些泵（分别用于缓冲液、生物分子溶液和沉淀剂溶液）由使用图像数据的控制算法控制。在第一阶段的研究，即，提供几微升的解决方案，以微反应室在几个小时内，并在这样的腔室中生长晶体的能力，电子测量的能力，以低于nL/s的流量与米比没有移动部件，并确定一个非常便宜的数字成像和分析方法。第二阶段研究的具体目标是（1）将这些新组件集成到单个自动化结晶系统中，以通过单个控制计算机测试所有功能的协调;（2）在（1）通过光刻设计和制造集成的4室阵列的基础上，包括光学器件、微流体器件和电子器件;以及（3）针对真实的晶体生长问题测试该阵列并改进基于图像的控制算法。该技术使用液-液结晶方法的自动化动力学控制作为一种手段，以检查化学条件的连续性，而不是使用大规模的离散筛选来提高这种相当昂贵的方法中的产量。由此产生的产品，一个结晶器微阵列（“微阵列”），将销售给世界上的大型制药公司，大学实验室与结构生物学单位和政府空间研究机构-一个市场，SHOT，Inc.很熟悉其实施。