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.
Thesaurus Terms: Caenorhabditis elegans, hybridoma, immunologic substance development /preparation, method development, monoclonal antibody, reagent /indicator helminthic antigen, high throughput technology biotechnology, enzyme linked immunosorbent assay, flow cytometry, laboratory mouse
