We generated a panel of monoclonal antibodies (MABs), specific for the enzyme elongation factor-1a (EF-1a), that are capable of inhibiting translation in Sf21 cell lysates. Three of the MABs (5H3, 7D6, and 7G3) preferentially recognize a 55 kD form of EF-1a, while a fourth antibody (MAB 8B7) most avidly binds to a 63 kD form. To determine whether MABs 5H3, 7D6, and 7G3 recognize the same or different epitopes, EF-1a was subjected to peptide mapping and immunoblot analysis. Peptide maps generated with trypsin yielded identical immunoblot patterns whether probed with MAB 5H3, 7D6, or 7G3; the same was true of peptide maps generated with V8 protease. After a 1-h incubation with trypsin, all three antibodies recognized EF-1a peptides ranging in mass from 25.5 to 55 kD; after 18 h, immunoreactive peptides of 49.9, 45.2, and 34.3 kD were still detected. Incubation of EF-1a with V8 protease for 1 h generated immunoreactive peptides ranging in mass from 18.3 to 55 kD; after overnight digestion, EF-1a peptides of 30.2 and 37.8 kD remained. From these limited findings, MABs 5H3, 7D6, and 7G3 appear to recognize EF-1a epitopes that are identical or spatially close together. Some EF-1a fragments recognized by MABs 5H3, 7D6, and 7G3 also contained epitopes reactive with MAB 8B7.
Keywords: translation, monoclonal antibodies
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