应用CE-SDS测定抗CD52人源化单克隆抗体参比品单体纯度及非糖化重链比例

目的:测定抗CD52人源单克隆抗体参比品单体纯度及非糖基化重链比例。方法:采用PA800 plus毛细管电泳系统,非还原十二烷基苯硫酸钠-毛细管电泳(CE-SDS)测定抗CD52人源单克隆抗体单体纯度,以及用还原CE-SDS电泳测定非糖化重链比例。结果:非还原CE-SDS三次测定单体纯度平均值为93.57%,主峰的迁移时间及修正峰面积百分比RSD分别为0.16%和0.19%;还原CE-SDS电泳重链修正峰面积百分比平均值为66.89%,修正峰面积百分比及迁移时间RSD分别为0.09%和0%;轻链修正峰面积百分比平均值为32.30%;轻链修正峰面积及迁移时间RSD分别为0%和0%;非糖基化重链修正峰面积百分比平均值为0.87%,修正峰面积百分比及迁移时间RSD分别为6.66%和0.27%。结论:CE-SDS测定抗CD52人源单抗单体纯度及非糖化重链比例实验结果偏差较小,表明结果准确可靠。

CE-SDS were Applied to Determine Monomer Purity and Proportion of Non-glycosylated Heavy Chain in Reference Products of anti-CD52 Humanized Monoclonal Antibody

ABSTRACT Objective: To determine the monomer purity and proportion of non-glycosylated heavy chain in reference productsanti- CD52 humanized monoclonal antibody. Methods: Using PA800 plus capillary electrophoresis system, Non-reduced CE-SDS were ap- plied to determine monomer purity and reduced CE-SDS were applied to determine non-glycosylated heavy chain in anti-CD52 human- ized monoclonal antibody. Results: Average value of monomer purity is 93.57% by Non-reduced CE-SDS in three times, RSD of migra- tion time of main peak and corrected area percent were 0.16% and 0.19% respectively. Average value of heavy chain corrected area per- cent is 66.89% by reduced CE-SDS in three times, RSD of migration time and corrected area percent of main peak were 0.09% and 0% respectively. Average value of light chain corrected area percent is 32.30% by reduced CE-SDS in three times, RSD of corrected area percent and migration time of main peak were 0% and 0% respectively. Average value of non-glycosylated heavy chain corrected area percent is 0.87% by reduced CE-SDS in three times, RSD of corrected area percent and migration time of main peak were 6.66% and 0.27% respectively. Conclusion: The data of using CE-SDS determine the monomer purity and proportion of non-glycosylated heavy chain in anti-CD52 humanized monoclonal antibody displayed a much small RSD, and manifested the result were accurate and reliable.