单克隆抗体与葡萄球菌肠毒素A结合物的制备及其抗肝癌作用

目的：制备超抗原（BAg)葡萄球菌肠毒素A（SEA）与抗人原发性肝细胞癌（简称肝癌）单克隆抗体（McAb)HAb18F(ab‘)2段的结合物，探讨其介导外周血单个核细胞（PBMC）的抗人肝癌作用。方法，制备McAb HAb18，并用木瓜蛋白酶消化法制备其F（ab‘)2段，再用化学连接剂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，结合物经经快速蛋白质液相层析系统用凝胶色谱柱纯化后，用SDSPAGE鉴定各种收集峰，免疫组化ABC法鉴定结合物的抗体活性，MTT法鉴定结合物活化PBMC的活性及其介导PBMC的杀伤肝癌细胞作用，结果，制备的HAb18F(ab‘)2-SEA结合物具有肝癌抗体活性和活化PBMC的功能，此结合物具有明显的介导PBMC杀伤肝癌细胞的作用，并与其浓度呈正相关，结论，SPDP是一种很好的蛋白质交联剂，HAb18 F(ab‘)2-SEA结合物民向治疗肝癌具有一定的发展前景。

Preparation of the Conjugate of Monoclonal Antibody and Staphylococcal Enterotoxin A and Its Anti-hepatoma Effect

Objective To prepare the conjugate of superantigen (SAg) Staphylococcal enterotoxin A (SEA) and monoclonal antibody (McAb) against human primary hepatocellular carcinoma (HCC) HAb18 F(ab') 2 fragment, and toprobe into the antiHCC effect of peripheral blood mononuclear cells (PBMC) targeted by HAb18 F(ab') 2 -SEA conjugate.Methods MAb HAb18 was extracted and its F(ab') 2 fragment was prepared by papainic digestion. The conjugate of HAb18 F(ab') 2 and SEA was prepared with chemical conjugater N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP).Every collected peak was identified by SDS-PAGE after purified with gel chromatography by fast protein liquid chromatography system. The antibody activity of the conjugate was identified by immunohistochemical ABC method. The activity of the conjugate to activate PBMC and the antiHCC effect of PBMC targeted by HAb18 F(ab') 2 -SEA was tested by MTT method.Results The prepared conjugate of HAb18 F(ab') 2 and SEA had the activity of antibody against HCC and activating PBMC. This conjugate had obvious effect of targeting PBMC to kill HCC cells, and its effect is correlated positively with the concentration of HAb18 F(ab') 2 -SEA.Conclusion SPDP is a kind of good protein conjugation. Targeting therapy of HCC with HAb18 F(ab') 2 -SEA conjugate has a hopeful prospect.