[Preliminary study of LFn-MAGE3 fusion protein expression and its biological function].

AIM: To explore the possibility of using the nontoxic form of anthrax toxin in cancer immunotherapy by LFn-MAGE3 fusion protein expression.

METHODS

A fusion expression vector named PET21a-LFn was constructed by inserting LFn coding senquence into PET21a. PET21a-LFn-MAGE3 fusion protein expression vector was constructed by cloning the whole MAGE-3 gene into plasmid PET21a-LFn. Q sepharose FF and Phe HP columns were employed to purify the fusion protein. The biological activity of LFn-MAGE3 was determined by cell test of repressing the cytotoxity of LF and the tests of immunofluscence of mouse macrophage.

RESULTS

The resulting plasmid expressed fusion protein LFn-MAGE3 in the soluble form in E.coli BL21, the cell tests showed that purified LFn-MAGE fusion protein was delivered into macrophage effectively with the help of PA(anthrax protective antigen).

CONCLUSION

The successful delivery of fusion protein into macrophages coordinated by PA may lay the foundation for its further use in cancer immunotherapy in animal experiments.