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Targeting therapy of single chain immunotoxin hdsFv-RC-RNase against hepatocellular carcinoma
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Abstract:
Objective: To observe the targeting therapy of the hdsFv-RC-RNase recombinant single chain immunotoxin on the xenograft of the human hepatocellular carcinoma in nude mice and to explore its clinical potentiality. Methods: The prokaryotic expression vector TIG-hdsFv-RC-RNase was transformed into E.coli BL21（DE3）plys to largely express recombinant single chain immunotoxin hdsFv-RC-RNase against hepatocellular carcinoma induced by IPTG. The expressed product was purified via Ni-NTA affinity chromatography under native conditions and mildly refolded. The ELISA was used to analyze its immunological activity of antigen-binding capability. The xenogrft model of the human hepatocellular carcinoma in nude mice was established and the targeting therapy of hdsFv-RC-RNase was evaluated. Results:After induced by IPTG, a new protein band with Mr 41 000 was found in the supernatant of the bacteria and expressed in a soluble form. The expressed product was purified to homogeneity via Ni-NTA affinity chromatography under native conditions. The results of ELISA showed the refolded hdsFv-RC-RNase had the specific antigen-binding capability to the human hepatocellular carcinoma cell, but not to the normal hepatocyte (P＜0.01). The targeting therapy on the xenograft in nude mice indicated that the efficiency of the hdsFv-RC-RNase was 100%(P＜0.01). The tumor inhibition rate reached 79.38%. Conclusion: The recombinant immunotoxin hdsFv-RC-RNase has been obtained sussefully, which has high activity and targeting therapy on hepatocellular carcinoma xinograft in nude mice. It may be used as targeting drug in therapy of hepatocellular carcinoma.
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