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Cytotoxity of EGF-PE35KDEL Chimeric Toxin on Hela Cell
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Abstract:
Objective: To construct a chimeric toxin consisting of a EGF linked to PE35KDEL and evaluate the potential of the EGF-PE35KDEL to target and kill tumor cells. Methods: By using of genetic engineering techniques, a recombinant expressing plasmid pET28a-EGF-PE35KDEL was constructed, the new obtained plasmid pET28a-EGF-PE35KDEL was transformed into E.coli BL21(DE3) for proposed protein expression. EGF-PE35KDEL was purified by DEAE-Sepharose FF chromatography, cytotoxity EGF-PE35KDEL on Hela cell growth was analyzed with crystal violet staining. Results: The recombinant plasmid pet28a-EGF-PE35KDEL was constructed and expressed in E.coli. The recombinant EGF-PE35KDEL protein was expressed as a soluble protein and was up to 20% of the total protein in E. coli BL21(DE3). With the two purification procedure, the EGF-PE35KDEL is about 95% pure. The IC50 of EGF-PE35KDEL on Hela cell is 0.07 μg/ml. Conclusion: Chimeric toxin EGF-PE35KDEL had cytotoxicity to Hela cells which over expressed EGFR in vitro.
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