【摘要】 抗p185erbB-2基因工程抗体是一种有潜力的抗肿瘤药物。以稳定表达抗p185嵌合抗体的重组工程CHO细胞株为对象,分别用不同浓度丁酸钠(0~2mmol/L)和丙酸钠(0~10mmol/L)对处在对数生长期的细胞进行处理,在连续5d的培养过程中,每隔24h取样测活细胞数量,并用ELISA检测上清中抗体含量,5d后结束培养用FACS检测细胞周期。同时还用丁酸钠和丙酸钠处理长至90%满度的细胞,然后每隔12h取样一次检测葡萄糖和乳酸的含量。结果表明丁酸钠和丙酸钠可以有效地提高嵌合抗体在工程CHO细胞中的表达,表达量最高时可达58.3~59.6mg/L,是对照组的1.5倍。同时抑制细胞生长和阻断细胞周期在G1期,并且可减少培养过程中葡萄糖的消耗和乳酸的生成。和丁酸钠相比,丙酸钠具有较小的细胞毒性,是一种有潜力的替代品。更多还原

【Abstract】 Overexpression of p185, which was encoded by the HER-2/erb2 proto-oncogene, has been reported in different types of tumors, especially in breast cancer. Athen recombinantengineering mouse/human chimeric antibody against p185 erbB-2, which showed the potential to inhibit proliferation of tumor cells overexpressing p185.A mouse/human chimeric antibody was constructed and expressed in the CHO-GS expression system. In the engineered CHO cells, the different concentrations of NaB (0.2,0.5,1-2mmol/L) or NaP (1,3,6-10mmol/L) was added respectively to the cultures of exponent growing cells in the presence of 5% serum. During 5 days cultivation, supernatants of the samples were taken every 24 hours for antibody detection by ELISA and the viable cell numbers were accounted using a hemocytometer. After 5 days, the cell cycles were analysed by the flow cytometry. In addition, NaB or NaP was added to the culture after the cells growing to close confluency and glucose consumption and lactate accumulation were measured every 12 hours. The results showed that both NaB and NaP addition can efficiently increase the antibody production and the highest chimeric concentration of yield was 58.3~59.6mg/L obtained by addition of either 1.0mmol/L NaB or 6.0mmol/L NaP, it is about 1.5 fold yield increase comparing to the control. NaB and NaP can also inhibit the cell growth and arrest the cells in at the G1 phase in a dose-dependent manner, but cell apoptosis was not observed during the cell culture, possibly due to the presence of serum. Although a similar increase in final chimeric antibody production on day 5 is obtained by either 1.0mmol/L NaB or 6.0mmol/L NaP, comparing with sodium butyrate, sodium propionate showed less influence on the cell growth and cyclecytotoxicity and could be a replacer of the sodium butyrate. In another test, NaB or NaP was added to the culture after the cells grew to near confluency and glucose consumption and lactate accumulation was measured every 12 hours. The results showed that the glucose consumption and lactate accumulation of CHO cells were reduced in the presence of NaB or NaP, which help to delay the cells decay and prolong the culture time.更多还原