Nanoliter Lab-on-a-chip for Protein Crystallization

描述（由申请人提供）：人类基因组含有至少30，000个独特的开放阅读框，其可产生> 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生物相关结构。为了获得单个蛋白质晶体，需要大量的蛋白质来筛选结晶的有利条件。蛋白质的3D结构需要了解任何有效的蛋白质工程，合理的药物设计和受控的药物输送。此外，许多令人感兴趣的蛋白质不幸地以有限的供应可用。 一个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，最需要的是占地面积小、完全自动化、成本降低以及能够在同一平台上控制大量参数。所有这些问题都直接通过开发芯片实验室的最新进展来解决。 在该项目的第一阶段，将开发一次性芯片实验室，用于蛋白质结晶筛选，建立在我们之前对纳米级水滴的电润湿操作的演示基础上，包括分配，运输，混合，分裂和稀释。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。微流体平台将被制造用于纳米级蛋白质、缓冲液和沉淀剂液滴的运输和分配。将在已知结晶条件下评价电润湿对溶菌酶、胰凝乳蛋白酶和GRP 94结晶的影响。将在集成芯片实验室平台上设置粗网格屏幕，用于演示软件可编程设置的筛选条件和溶菌酶结晶。

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.