[关键词]
[摘要]
目的:制备超抗原(SAg)葡萄球菌汤毒素A(SEA)与搞人肝细胞癌单克隆抗体(McAb)F(ab’)_2段的结合物,并观察其介导外周皿单个核细胞(PBAMC)的抗人肝癌作用,方法:提取McAb HAbl8,用木瓜蛋日酶制取其F(ab’)_2段,再用N-琥珀酰亚氨基-3-(2二硫吡啶)丙酸酯(SPDP)制备HAb18 F(ab’)_2-SEA结合物;经凝胶色谱柱纯化后用SDS-PAGE鉴定各收集峰,免疫组化ABC法鉴定此结合物的抗体活性,MTT法观察HAb18 F(ab’)_2SEA介导PBMC的抗肝癌作用.结果:成功地制备了HAb18 F(ab’)_2-SEA结合物,并且此结合物具有明显的介导PBMC靶向杀伤肝癌细胞的作用,并与其剂量呈正相关.对照组无此作用结论:MCAb与SAg结合物导向治疗肝癌是一种有发表前景的肝癌免疫治疗模式.
[Key word]
[Abstract]
Objective: To prepare the conjugate of supcrantigen (SAg) staphylococcal enterotoxin A (SEA) and monoclonal antibody (McAb) against human hepatocellular carcinoma HAbl8 F(ab' )_(2) fragment and to investigate the anti-human hepatoma effect of peripheral blood mononuclear cells (PBMC) targeted by HAbl8 F(ab' )i-SEA. Methods: McAb HAbl8 was extracted and its F(ab' )_(2) fragment was prepared with papain; the conjugate HAblS F(ab' )_(2)-SEA was prepared with N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP); eveny collected peak after purification was identified with gel chromatography; the activity of antibody in the conjugate was identified with immunohistocheinical ABC method; the anti-hepatoma effect of PBMC targeted by HAbl8 F(ab' )_(2)-SEA was observed with MTT method. Results: The conjugate HAbl8 F(ab' )_(2)-SEA was prepared successfully and it had obvious effect of targeting PBMC to kill hepatoma cells, and this effect is correlated positively with the dose of HAbl8 F(ab')_(2)-SEA. Control groups had no such effect. Conclusion: Targeting therapy of hepatoma with McAb-SAg conjugate might be a kind of hopeful model of hepatoma im-munotherapy.
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[基金项目]
国家863计划(102-09-01-02),国家自然科学基金(39770827)资助