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单克隆抗体博来霉素A6偶联物对白血病细胞特异性结合与内化
引用本文:田佩玉,张曼丽,黄静,于滨,甄永苏. 单克隆抗体博来霉素A6偶联物对白血病细胞特异性结合与内化[J]. 药学学报, 1989, 24(1): 16-21
作者姓名:田佩玉  张曼丽  黄静  于滨  甄永苏
作者单位:中国医学科学院医药生物技术研究所,中国药品生物制品检定所,中国医学科学院医药生物技术研究所,中国医学科学院医药生物技术研究所,中国医学科学院医药生物技术研究所 北京 100050,北京 100050,北京 100050,北京 100050
摘    要:
抗CCT2单克隆抗体博来霉素A6偶联物可吸附胶体金颗粒(McAb-A6-Au)。电镜观察表明,在4℃,1h,表面有McAb-A6-Au颗粒的CEM细胞最高达78%;在37℃,4h,内化McAb-A6-Au颗粒的CEM细胞高达72%。而抗原性无关的U937细胞仅为14%。并且McAb-A6-Au颗粒能直接穿过细胞膜、核膜进入细胞浆和细胞核。37℃,1h已有10~18%的CEM细胞核内有McAb-A 6-Au颗粒。实验结果提示了单抗与博来霉素A6的偶联物与选择性地结合靶细胞,而且进入细胞速度快、穿透力强,有可能成为治疗白血病药物。

关 键 词:单克隆抗体  博来霉素A 6  免疫结合物  胶体金  免疫电镜
收稿时间:1988-01-29

SPECIFIC BINDING AND INTERNALIZATION OF ANTI-CCT2 MONOCLONAL ANTIBODY AND BLEOMYCIN A6 CONJUGATE IN HUMAN LEUKEMIA CELLS
PY TIAN,ML ZHANG,J HUANG,B YU and YS ZHEN. SPECIFIC BINDING AND INTERNALIZATION OF ANTI-CCT2 MONOCLONAL ANTIBODY AND BLEOMYCIN A6 CONJUGATE IN HUMAN LEUKEMIA CELLS[J]. Acta pharmaceutica Sinica, 1989, 24(1): 16-21
Authors:PY TIAN  ML ZHANG  J HUANG  B YU  YS ZHEN
Abstract:
The immunoconjugate of anti-CCT2 monoclonal antibody linked to bleomycin A6 was adsorbed on colloidal gold particles (McAb-A6-Au). Binding and internalization of McAb-A6-Au particles in human leukemia CEM cells were examined by electron microscopy. After 60 min at 4 degrees C, McAb-A6-Au particles were bound to the surface membrane of 78% of CEM cells. Transferring to 37 degrees C for 15 min, McAb-A6-Au particles were found to be 56% inside the CEM cells and about one third of the cells contained particles in the nucleus. After 4 h at 37 degrees C the percentage of CEM cells containing McAb-A6-Au particles increased to 72%. However, only 14% of the antigenically irrelevant U937 cells contained these particles and none of them was found in the nucleus. Preincubation with unconjugated anti-CCT2 monoclonal antibody markedly blocked the McAb-A6-Au particle uptake in CEM cells. The McAb-A6-Au particles were internalized through the formation of endocytotic vesicles. In addition, some McAb-A6-Au particles were able to penetrate the plasma membrane directly into cytoplasma and notably into the nucleus. Results indicate that the immunoconjugate of monoclonal antibody linked to bleomycin A6 showed selective binding to target cells and entered the cells specifically and rapidly.
Keywords:Monoclonal antibody  Bleomycin A6  Immunoconjugate  Colloidal gold  Immunoelectron microscopy
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