抗癌抗生素力达霉素与抗IV型胶原酶单链抗体的基因工程组装融合蛋白

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目的 以IV型胶原酶为靶点 ,研制兼具有抗肿瘤侵袭转移和强烈杀伤肿瘤细胞的新型单抗导向药物。方法 力达霉素 (LDM)分子由力达蛋白 (LDP)和力达发色团 (LDC)两部分组成。从基因工程转化菌中提取融合蛋白 (LDP Fv)并与LDC组装重建 ,制备高效抗癌抗生素力达霉素与单链抗体的组装融合蛋白 (LDM Fv)。通过ELISA、酶谱分析、Boyden小室体外侵袭实验及克隆形成测定 ,观察其生物学活性。结果 LDM Fv能特异性与肿瘤细胞的IV型胶原酶结合并抑制其活性 ,LDP Fv有阻断肿瘤细胞侵袭的能力 ,但无细胞毒性 ,加入LDC制成的组装融合蛋白LDM Fv对肿瘤细胞有强烈的杀伤作用 ,并明显强于丝裂霉素和阿霉素。结论 LDM Fv可能成为一种新型高效、小型化抗肿瘤导向药物 Objective To develop novel monoclonal antibody-directed drugs with anti-tumor invasion and metastasis and strong killing of tumor cells using collagen type IV as a target. Methods The Lidamycin (LDM) molecule consists of two components, the LDP and the Lida chromophore (LDC). The fusion protein (LDP Fv) was extracted from genetically engineered transformants and reconstructed with LDC to prepare an assembled fusion protein (LDM Fv) of high-potency anticancer antibiotics, lidamycin and single-chain antibodies. The biological activity was observed by ELISA, zymogram analysis, Boyden chamber in vitro invasion assay and colony formation assay. Results LDM Fv specifically binds to type IV collagenase of tumor cells and inhibits its activity. LDP Fv has the ability to block tumor cell invasion, but it has no cytotoxicity. LDM Fv, an assembled fusion protein made from LDC, has been shown to have tumor cells. Strong killing effect, and significantly stronger than mitomycin and doxorubicin. Conclusion LDM Fv may become a novel, highly effective and miniaturized antitumor drug
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