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Abstract Epidermal Growth Factor Receptor is becoming a perfect target to kill carcinoma cells specially because of its overexpression on the surface of carcinoma cells. Cationic antimicrobial peptides (CAP) have its own special membrane-lytic cytotoxicity mechanism. In this study, the membrane-lytic immunotoxin (IT), named as chimeric protein MEGFMEL, which composed of mouse epidermal growth factor (MEGF) as targeting part and melittin (MEL) as cytotoxic part, was constructed to kill carcinoma cells with epidermal growth factor receptor (EGFR) overexpression. Using E.coli BL21 as expression strain and pET30a as expression vector, through low-temperature inducing expression and thawing & freezing purification without cytolysis procedure, the MEGFMEL obtained was of 63.45μg/mL concentration and 68% purity. In vitro activity measurement showed the interested MEGFMEL had significant killing effect to A431 cell of squamous epithelium carcinoma which overexpresses EGFR on its surface, with LD50 value 52.6μg/mL.
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Received: 20 September 2007
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