Expression of Anti-ASGPR ScFv Targeting Melittin and Its Anti-tumor Efficacy in Vitro

Objective To explore the cytolytic efficacy on liver cancercell line HepG_2 of the recombinant protein C1M formed by Melittin fusing to an anti-ASGPR single-chain Fv antibody(C1). Methods A single colony of E.coli XL1-Blue containing plasmid pGC-C1M was inoculated in LB broth,then diluted 1/100 into 1 000 mL LB broth and induced with 1(mmol·L~(-1)) IPTG.The recombinant C1M was purified with Ni~(2+) chelating HiTrap HP column.Its antigen-binding ability was evaluated with immnunohistochemistry,and growth inhibitor rate of tumor was analysed by MTT. Results SDS-PAGE showed that the purified recombinant protein C1M was expressed in E.coli as soluble style with a molecular weight of about 29.4×10~3.The result of immnunohistochemistry showed that C1M could effectively recognized ASGPR.On the third day of culturing HepG2 with C1M,the cellular growth inhibiting rate reached 92.0%. Conclusion Successful expression of C1M was achieved in(E.coli),and Melittin at its C end can effectively inhibit the growth of tumor cells.