[
International C. elegans Meeting,
2001]
To construct antibody libraries covering C elegans whole proteins, we have been in an effort to produce monoclonal antibodies by the methods of cell fusion and phage display. As a source of these antibodies supply, we chose the spleen cells of Balb/c mouse immunized with homogenates of C elegans whole body or with proteins derived from C elegans cDNA expression library supplied by A.Fire. In a preliminary work, we tried to produce monoclonal antibodies by the fusion of P3U1 myeloma cells with spleen cells from the mice immunized with C elegans whole proteins mixed with Freunds adjuvant. We obtained the antisera showing strong reaction to many kinds of C elegans proteins, and could produce 16 kinds of monoclonal anti- C elegans antibodies named 4A7, 4A3, 4A1, 1E1, 2G5, 4A9, 1B1, 4C10, 2B8, 2B6, 3B12, 2H3, 3A6, 1F6, 1D12 and 1C10. When we immunized the mice with C elegans whole proteins alone instead of mixture with Freunds adjuvant, we obtained antisera showing weak reaction to C elegans proteins and could produce only one kind of monoclonal anti- C elegans antibody named 3D7. We applied these antisera and monoclonal antibodies on western blotting analysis to identify their antigens. The antisera reacted with many kinds of C elegans proteins. On the other hand, monoclonal antibody 4A7, 4A1, 1E1, 2G5, 2B6, 3B12 and 2H3 reacted with 40kDa C elegans protein alone suggesting these antibodies detecting a single molecule. In the same way, 55kDa protein was suggsted as a sole antigen of 3A6, 1F6 and 1D12. A number of antigens were suggested for 4A3, 4A9, 1B1, 4C10, 2B8 and 3D7. The immunohistochemical localization of these antigens in C elegans is under investigation.