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Construction and Expression of the EGF-Ang Fusion Protein and Its Biological Activity
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Abstract:
Objective：To construct a humanized fusion protein consisting of a EGF linked to a human angiogenin(Ang) and evaluate the potential of the construct(EGF-Ang) to target and kill tumor cells.Methods：By using of genetic engineering techniques, we constructed a recombinant expressing plasmid pEGF-Ang, and expressed fusion protein accumulated in intracellular inclusion bodies. The recombinant EGF-Ang proteins were purified through DEAE-Sepharose FF chromatography in the denatured condition. After renaturing process, the cytotoxicity was measured with MTT colorimetric assay in vitro. Results：A novel fusion protein named EGF-Ang was constructed and expressed in E coli. Analysis of SDS-PAGE and thin layer scanning results showed that amount of expressed fusion protein was 18.6% in total lysis protein of bacteria. Cytotoxicity analysis showed that the fusion proteins could inhibit the growth of Hep2 cells but had little influence on the growth of MA104 cells. Conclusion：EGF-Ang fusion protein had definitely cytotoxicity to Hep2 cells which over expressed EGFR in vitro.
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