Elongation factor-1a (EF-1a) is an enzyme necessary for protein translation. It is responsible for the binding of aminoacyl tRNA to the acceptor A site on the ribosome during eukaryotic protein chain elongation. Because it is an essential enzyme, EF-1a offers an excellent target for the disruption of insect metabolism, but agents known to interfere with EF-1a activity are toxic to humans. During recent months, our goal has been to develop monoclonal antibodies (Mabs) that can disrupt EF-1a activity in insects without cross-reacting with the human enzyme. Mabs were generated to EF-1a derived from Sf21 cells by immunizing mice with the enzyme eluted from SDS-PAGE gels. The Mabs were screened for cross-reactivity to human EF-1a by immunoblotting against a human carcinoma cell line. The Mabs were further screened for the ability to recognize EF-1a in its native conformation by ELISA and immunoprecipitation assays that employed cell lysates or chromatographically purified EF-1a as the antigen. Mabs that recognized the native enzyme in insects, but not humans, were added to lysates of Sf21 cells to determine whether any of the antibodies could inhibit incorporation of 35S-methionine into newly-synthesized in vitro translation products. Of the four EF-1a-specific Mabs tested, two significantly inhibited protein synthesis when added to Sf21 lysates at a 1:1 antibody-to-enzyme molar ratio (P<0.05).
Species 1: Lepidoptera Noctuidae Spodoptera frugiperda (fall armyworm)
Keywords: elongation factor-1 alpha, protein synthesis
The ESA 2001 Annual Meeting - 2001: An Entomological Odyssey of ESA