Monoclonal antibodies to elongation factor-1α inhibit in vitro translation in lysates of Sf21 cells

Document Type

Article

Publication Title

Archives of Insect Biochemistry and Physiology

Abstract

Elongation factor-1α (EF-1α) is an enzyme that is essential for protein synthesis. Although EF-1α offers an excellent target for the disruption of insect metabolism, agents known to interfere with EF-1α activity are toxic to humans. In this article, we describe the development of monoclonal antibodies (MAbs) that can disrupt the activity of insect EF-1α without cross-reacting with the human enzyme. MAbs were generated to EF-1α from Sf21 cells derived from the fall ormyworm, Spodoptera frugiperda, by immunizing mice with EF-1α eluted from SDS-PAGE gels. The MAbs reacted with EF-1α in eggs and first through fifth instars of the fall armywarm in immunoblots of SDS-PAGE gels, but did not recognize EF-1α in human carcinoma cells and normal tissues. MAbs with the ability to recognize EF-1α in its native conformation, identified through immunoprecipitation experiments, were added to Sf21 cell lysates to determine whether the antibodies could inhibit incorporation of [35S]methionine into newly synthesized in vitro translation products. Of the four EF-1α-specific MAbs tested, three significantly inhibited protein synthesis when compared to the negative control antibody (P < 0.001, one-way ANOVA; followed by Dunnett's test, P < 0.05). © 2003 Wiley-Liss, Inc.

First Page

17

Last Page

34

DOI

10.1002/arch.10061

Publication Date

1-1-2003

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