人鼠嵌合单克隆抗体c30.6的纯化

Purification of a chimeric monoclonal antibody c30.6

采用介体电泳技术对CHO细胞培养液中的人鼠嵌合单克隆抗体c30 6进行纯化 ,第一步在pH8 6条件下进行 ,抗体带正电荷而培养液中的大部分杂质带负电荷 ,抗体和杂质在电场作用下向两电极运动 ,结果被分离在分离膜的两侧 ,SDS -PAGE显示抗体纯度达 95 % ,且剩余杂质的分子量均小于抗体的分子量。第二步是在pH6 0条件下进行 ,此时抗体和杂质都带正电荷 ,它们在电场作用下向同一方向运动 ,但分离膜只允许分子量较小的杂质通过 ,抗体得以进一步纯化 ,抗体的回收率为 80 % ,而且生物活性未受影响。

The preparative electrophoresis was used to purify a mouse/human IgG1 chimeric antibody c30 6 from CHO cell culture supernatant The chimeric monoclonal antibody carried a positive charge at pH 8 6,whereas most contaminating host-cell proteins carry a negative charge This property was used to draw contaminating proteins through a separation membrane towards the anode of the separation system,whilst the antibody remained at the cathode By SDS-PAGE,the purity of the antibody solution was qualitatively estimated to be >95% pure and the molecular weight of residual impurities was less than that of antibody At second step,both antibody and impurities carried positive charge in pH6 0,but residual contaminant was removed by crossing though the separating membrane, whilst antibody stayed The recovery of antibody was over 80% and the antibodies have activity