Novel Caenorhabditis Elegans Reagents

描述(申请人提供)：线虫被公认为是一种非常有用的模式生物。第十四届国际线虫年会于2003年6月29日至7月3日举行，发表了2500多名作者的1151多篇摘要。在20世纪80年代，对线虫胚胎细胞谱系和成年神经系统连接的完整描述发表了。1998年公布了一个基本完整的基因组序列，激发了许多大规模研究，整个科学界都受益于由此产生的工具。例如，图谱芯片阐明了协调的基因表达，RNA干扰(RNAi)研究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线虫蛋白质组奠定了基础。这将需要研究蛋白质的表达、细胞定位和功能。人们普遍认为，特定的抗体是这类工作不可缺少的工具，我们相信，单抗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个预测线虫基因产物的cDNA已经被鉴定。然而，只有大约100种针对线虫抗原的抗体可以从商业或其他收费的服务库获得。创造特定的抗体是昂贵的，耗时的，并且充满了许多技术问题。Abeome Inc.开发了一种高通量、基于杂交的平台，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体。我们已经开发了一种制备杂交瘤的方法，该杂交瘤能够旺盛地分泌并表面呈现Ig。这一创新消除了限制稀释克隆的劳动密集型和问题缠身的步骤，因为这些杂交瘤可以通过荧光激活细胞分选(FAC)来选择，培养可以自动化。这个第一阶段的项目将是申请者和史蒂文·L博士在埃默里大学的埃尔诺实验室之间的合作努力。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L博士的Hernault博士鉴定的五种线虫蛋白的抗体。申请者和L博士的Hernault将在免疫原设计、抗体生产和表征方面进行合作。预计在第一阶段工作中获得的数据将支持在第二阶段项目中制备数百或数千种线虫试剂的可行性。

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.