Novel Caenorhabditis Elegans Reagents

新型线虫试剂

• 批准号: 6789205
• 负责人: PAUL W PRICE
• 金额: $ 10万
• 依托单位: ABEOME CORPORATION
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-03 至 2004-11-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6789205
• 关键词: Caenorhabditis elegans; biotechnology; enzyme linked immunosorbent assay; flow cytometry; helminthic antigen; high throughput technology; hybridomas; immunologic substance development /preparation; laboratory mouse; method development; monoclonal antibody; reagent /indicator

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.

描述（由申请人提供）：秀丽隐杆线虫被认为是一种非常有用的模式生物。 2003 年 6 月 29 日至 7 月 3 日举行的第 14 届国际线虫双年度会议征集了 2500 多名作者发表的 1151 多篇摘要。 20 世纪 80 年代，发表了关于秀丽隐杆线虫胚胎细胞谱系和成体神经系统布线的完整描述。 1998 年出版的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，而RNA干扰（RNAi）研究已经允许敲低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为全面了解秀丽隐杆线虫蛋白质组奠定了基础。这需要对蛋白质表达、细胞定位和功能进行研究。人们普遍认为，特异性抗体是此类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佳解决方案，至少 17,298 个预测的秀丽隐杆线虫基因产物的 cDNA 已被鉴定。然而，只有大约 100 种针对线虫抗原的抗体可从商业或其他收费服务存储库中获得。制造特异性抗体既昂贵又耗时，并且充满许多技术问题。 Abeome Inc. 开发了一种基于杂交的高通量平台，能够以前所未有的速度生产单克隆抗体，并且成本远低于任何其他方法。我们开发了一种制备杂交瘤的方法，该杂交瘤能够强劲地分泌和表面呈递 Ig。这项创新消除了有限稀释克隆的劳动密集型和问题多的步骤，因为这些杂交瘤可以通过荧光激活细胞分选（FACS）进行选择，并且平板接种可以自动化。该第一阶段项目将由申请人和埃默里大学 Steven L'Hernault 博士的实验室合作完成。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'Hernault 博士确定的五种秀丽隐杆线虫蛋白具有反应性的抗体。 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。