Nanoliter Lab-on-a-chip for Protein Crystallization

用于蛋白质结晶的纳升芯片实验室

• 批准号: 6834059
• 负责人: VAMSEE K. PAMULA
• 金额: $ 14.96万
• 依托单位: ADVANCED LIQUID LOGIC
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6834059
• 关键词: biomedical equipment development; chymotrypsin; computer program /software; consumable /disposable biomedical equipment; crystallization; electrodes; fluid flow; gastrin releasing peptide; lysozyme; nanotechnology; protein engineering; protein structure; protein transport

DESCRIPTION (provided by applicant): The human genome contains at least 30,000 unique open reading frames that may yield >100,000 polypeptide products. These products assemble into more than a million biologically relevant structures of interest per organism. In order to obtain a single protein crystal, large amounts of that protein are required to screen for the favorable conditions for crystallization. The 3D structure of proteins needs to be understood for any effective protein engineering, rational drug design, and controlled drug delivery. Moreover, many interesting proteins are unfortunately available in limited supply. A protein crystallization screening platform that consumes only nanoliters of protein solutions will help reduce the consumption of the precious proteins. In addition, small foot print area, complete automation, reduced cost, and the ability to control a large number of parameters on the same platform would be most desirable. All these issues are directly addressed by the recent advances in developing a lab-on-a-chip. In phase I of this project, a disposable lab-on-a-chip will be developed for protein crystallization screening building on our previous demonstrations of electrowetting manipulation of nanoliter-sized water droplets including dispensing, transport, mixing, splitting, and dilution. Phase I work will particularly focus on demonstrating the feasibility of an electrowetting-based lab-on-a-chip for protein crystallization screening. A microfluidic platform will be fabricated for transport and dispensing of nanoliter-sized protein, buffer, & precipitant droplets. The effect of electrowetting will be evaluated on the crystallization of Lysozyme, Chymotrypsin, and GRP 94 under known crystallization conditions. A coarse grid screen will be setup on the integrated lab-on-a-chip platform for demonstrating software programmable set up of screening conditions and crystallization of Lysozyme.

描述（由申请人提供）：人类基因组包含至少 30,000 个独特的开放阅读框，可产生 >100,000 种多肽产物。这些产品组装成每个生物体超过一百万个感兴趣的生物学相关结构。为了获得单一蛋白质晶体，需要大量该蛋白质来筛选有利的结晶条件。任何有效的蛋白质工程、合理的药物设计和受控药物输送都需要了解蛋白质的 3D 结构。此外，不幸的是，许多有趣的蛋白质供应有限。 仅消耗纳升蛋白质溶液的蛋白质结晶筛选平台将有助于减少珍贵蛋白质的消耗。此外，占地面积小、完全自动化、成本降低以及在同一平台上控制大量参数的能力将是最理想的。所有这些问题都可以通过芯片实验室开发的最新进展直接解决。 在该项目的第一阶段，将基于我们之前对纳升大小水滴的电润湿操作（包括分配、运输、混合、分裂和稀释）的演示，开发用于蛋白质结晶筛选的一次性芯片实验室。第一阶段的工作将特别侧重于证明基于电润湿的芯片实验室用于蛋白质结晶筛选的可行性。将制造一个微流体平台，用于运输和分配纳升大小的蛋白质、缓冲液和沉淀剂液滴。将在已知的结晶条件下评估电润湿对溶菌酶、胰凝乳蛋白酶和GRP 94的结晶的影响。将在集成芯片实验室平台上设置粗网格屏幕，用于演示筛选条件和溶菌酶结晶的软件可编程设置。