Novel Caenorhabditis Elegans Reagents

描述（由申请人提供）：C。秀丽线虫是一种非常有用的模式生物。第14届双年国际C。2003年6月29日至7月3日举行的秀丽隐杆线虫会议吸引了2500多位作者发表的1151篇摘要。在20世纪80年代，对C. elegans出版了。1998年基本完整的基因组序列的发表激发了许多大规模研究，整个科学界都从由此产生的工具中受益。例如，分析微阵列已经阐明了协调的基因表达，并且RNA干扰（RNAi）研究已经允许敲除许多基因以揭示功能表型的减少。这些和其他广泛的先前研究已经为全面了解C。线虫蛋白质组这将需要研究蛋白质表达、细胞定位和功能。人们普遍认为特异性抗体是这种努力不可或缺的工具，我们相信单克隆抗体为所需的大规模完整蛋白质组分析提供了最好的解决方案，至少17，298个预测的C.已经鉴定了秀丽线虫的基因产物。然而，只有大约100种抗体针对C。线虫抗原可从商业或其他收费服务库获得。产生特异性抗体是昂贵的，耗时的，充满了许多技术问题。Abeome Inc.已经开发出一种高通量的基于杂交瘤的平台，用于以前所未有的速度和比任何其他方法低得多的成本生产单克隆抗体。我们已经开发了一种制备稳健分泌和表面呈递IG的杂交瘤的方法。这种创新消除了劳动密集型和问题困扰的有限稀释克隆步骤，因为这些杂交瘤可以通过荧光激活细胞分选（FACS）进行选择，并且可以自动进行平板接种。这个第一阶段的项目将是申请人和埃默里大学Steven L'Hernault博士实验室之间的合作。我们将使用传统的杂交瘤方法和我们的创新方法来制备针对五种C。L'Hernault博士发现的蛋白质。 申请人和L'Hernault博士将在免疫原设计、抗体生产和表征方面进行合作。预计在第1阶段工作中获得的数据将支持制备数百或数千个C。elegans试剂在第二阶段的项目。

DESCRIPTION (provided by applicant): C. elegans is well established as a highly useful model organism. The 14th annual Biennial International C. elegans Conference held June 29th-July 3, 2003 elicited over 1151 abstracts published by over 2500 authors. In the 1980s, complete descriptions of the embryonic cell lineage and the adult nervous system wiring for C. elegans were published. Publication of an essentially complete genomic sequence in 1998 inspired numerous large-scale studies, and the entire scientific community has benefited from the resulting tools. For example, profiling microarrays have elucidated coordinated gene expression and RNA interference (RNAi) studies have allowed knockdown of many genes to reveal reduction of function phenotypes. These and other extensive prior studies have set the stage for a complete understanding of the C. elegans proteome. This will require studies of protein expression, cellular localization and function. It is universally recognized that specific antibodies are indispensable tools for such efforts and we believe monoclonal antibodies offer the best solution for the required large scale of a complete proteome analysis, cDNAs for at least 17,298 predicted C. elegans gene products have been identified. Yet there are only about 100 antibodies directed against C. elegans antigens available from commercial or other fee for service repositories. Creating specific antibodies is expensive, time consuming and fraught with many technical problems. Abeome Inc. has developed a high-throughput, hybridorna-based platform for producing monoclonals with unprecedented speed and at substantially lower costs than by any other method. We have developed a method of preparing hybridomas that robustly secrete and surface present Ig. This innovation eliminates the labor intensive and problem-plagued step of limit dilution cloning because these hybridomas can be selected by Fluorescent Activated Cell Sorting (FACS) and plating can be automated. This phase 1 project will be a collaborative effort between applicant and Dr. Steven L'Hernault's lab at the Emory University. We will use both conventional hybridoma methods and our innovative methods to prepare antibodies reactive against five C. elegans proteins identified by Dr. L'Hernault. Applicant and Dr. L'Hernault will collaborate on immunogen design, antibody production and characterization. It is expected that data obtained in this Phase 1 effort will support the feasibility of preparing hundreds or thousands of C. elegans reagents in a Phase 2 project.