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，可能产生100万到10万种多肽产物。这些产品组装成每个生物体感兴趣的超过一百万个生物相关结构。为了获得一个单一的蛋白质晶体，需要大量的蛋白质来筛选结晶的有利条件。为了进行有效的蛋白质工程、合理的药物设计和控制药物递送，需要了解蛋白质的三维结构。此外，不幸的是，许多有趣的蛋白质供应有限。