人鼠嵌合Fab抗体通用表达载体的构建和抗人肝癌相关抗原HAb18G嵌合Fab抗体的表达

图1 重组通用表达载体pComb3C的限制性酶切分析. M: DNA marker; 1: Human IgG3CH1; 2: Constant region of human κchain; 3: pComb3C/Xho I + Spe I; 4: pComb3C/Sac I+Xba I; 5: pComb3C/EcoR Ⅴ+Sal I.

图2 重组cFab表达载体pComb3C/cFab-gIII的限制性酶切分析. M: DNA marker; 1: VH; 2: VL; 3: pComb3C/cFab-gIII/ Xho I + Spe I; 4: pComb3C/cFab-gIII/ EcoRⅤ+ Spe I; 5: pComb3C/cFab-gIII/Sac I + Xba I; 6: pComb3C/cFab-gIII/ Sal I + Xba I; 7: pComb3C/cFab-gIII/ Spe I+Nhe I.

图3 cFab基因表达载体的构建.

图4 夹心ELISA检测cFab的表达. 1: pComb3 after induction; 2: pComb3C/cFab before induction; 3: pComb3C/cFab after induction; 4: chHAb18; 5: PBS.

图5 夹心ELISA检测cFab抗体表达产物的定位. 1: PBS; 2: Media supernatant; 3: Periplasmic extracts; 4: Cytoplasmic extracts; 5: Precipatation.

图6 cFab抗体表达产物的SDS-PAGE电泳和Western-Blot分析. 1: mM marker; 2: Reduced cFab; 3: Unreduced cFab; A: Western blot of reduced cFab; B: Western blot of unreduced cFab.

图7 间接ELISA检测cFab抗体的抗原结合特异性.

图8 竞争性ELISA检测cFab与HAb18的相对抗原结合活性.

图9 免疫荧光染色观察cFab与肝癌细胞株HHCC的结合(×200). A: cFab; B: human IgG.