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目的制备符合人用标准的抗人肝癌mAbHAb18F(ab′)2 片段。方法用胃蛋白酶消化经硫酸铵盐析的腹水抗体 ,然后在快速蛋白液相色谱(FPLC)上用Phenyl SepharoseHP疏水柱纯化并进行质检。结果经Phenyl SepharoseHP疏水柱纯化后 ,F(ab′)2 片段的纯度可达98.8% ;回收率大于50 % ;免疫活性为1∶8000;细菌、热原质检测均呈阴性。结论本制备工艺周期短、易于质控 ,适宜进行中试放大及半成品生产。
Objective To prepare a human anti-human hepatoma mAb HAb18F (ab ’) 2 fragment that is suitable for human use. Methods Ascites antibodies were salted by ammonium sulfate with pepsin and then purified on Phenyl Sepharose HP Hydrophobic Columns on a Fast Protein Liquid Chromatography (FPLC) and checked for quality. Results The purity of F (ab ’) 2 fragment was 98.8% after purification by Phenyl Sepharose HP column. The recovery was more than 50% and the immunogenicity was 1: 8000. The bacterial and pyrogen detection were all negative. Conclusion The preparation process is short, easy to control, suitable for enlargement and semi-finished pilot production.