三氧化二砷免疫毫微球的制备及活性检测

Preparation of arsenic trioxide-loaded albumin immuno-nanospheres and detection of its on bladder tumor in vitro

目的 制备单抗BDI 1导向的三氧化二砷免疫蛋白微球 [As2 O3 (BSA NS) BDI 1]并检测其特异性结合膀胱肿瘤细胞的活性。方法 通过蛋白交联固化及N 羟基琥珀酰亚胺基 3 (2 吡基二硫 ) 丙酸脂 (SPDP)交联的方法制备蛋白免疫微球。还原电泳、光镜和电镜的方法鉴定As2 O3 免疫微球的共价连接与活性。吖啶橙染色检测肿瘤细胞凋亡。结果 还原电泳可见免疫微球被还原后在远端形成两条蛋白条带。光镜下可见蛋白微球在肿瘤细胞周围并随细胞移动。电镜下可见蛋白微球与肿瘤细胞紧密连接且形态完整。吖啶橙染色可见免疫微球处理的膀胱肿瘤细胞表现出凋亡特征。结论 制备的As2 O3 (BSA NS) BDI 1由共价键连接且能特异性的结合膀胱肿瘤细胞BIU 87而发挥诱导凋亡的作用。

Objective To preparare arsenic trioxide-loaded albumin Immuno-nanospheres As_2O_3-(BSA-NS)-BDI-1 targeted with Monoclonal Antibody BDI-1 and test its specific killing effect on bladder tumor cell.Methods Immuno-nanospheres were prepared by methods of protein cross-linking solidify and SPDP cross-linking.Immuno-nanospheres and its activity were tested by reduction electrophoresis,microscopy and scanning electron microscopy.The acridine orange staining methods were used to assay the apoptosis of tumor cells in vitro.Results Immuno-nanospheres which were reduced with DTT showed two protein bands at distant distal end.Under microscopy,the BIU-87 cells were rounded with immuno-nanospheres and As_2O_3-(BSA-NS)-BDI-1 moved with the BIU-87 cells.The scanning electron microscopy revealed the albumin immuno-nanospheres were tight junctioned with the BIU-87 cells.The acridine orange staining showed apoptosis of BIU-87 cells.Conclusion As_2O_3- (BSA-NS)-BDI-1 might specifically bind against BIU-87 cells by covalent bond and induce BIU-87 cells apoptosis.