郭秋菊;李雪;封纯芳;李乐;陈红霞;于玉根;

• 1:深圳万乐药业有限公司抗体中心

摘要(Abstract)：

目的比较抗人类表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)单克隆抗体酸性电荷异构体、原液及纯化工艺中阳离子交换层析预洗脱杂质质量差异,为细胞培养工艺研究和纯化工艺参数的调整提供参考。方法采用体积排阻色谱法(size exclusion chromatography,SEC)检测供试品单体、聚体和残体含量;非还原/还原毛细管电泳-十二烷基磺酸钠(capillary electrophoresis-sodium dodecyl sulfate,CE-SDS)法检测供试品电泳纯度;弱阳离子交换色谱法(weak cation exchange chromatography,CEX)检测供试品电荷异构体分布;毛细管等电聚焦(capillary iso-electric focusing,c IEF)法测定供试品等电点;细胞增殖抑制法测定供试品生物学活性。结果 SEC检测结果显示,预洗脱杂质、酸性异构体和原液的单体含量分别为54.44%、99.08%和99.80%,残体含量分别为44.33%、0.61%和0。非还原CE-SDS结果显示,预洗脱杂质、酸性异构体和原液的主峰含量分别为53.93%、90.83%和95.83%,小分子杂质含量分别为46.07%、9.17%和4.17%;还原CE-SDS结果显示,预洗脱杂质、酸性异构体和原液的轻、重链百分含量分别为78.00%、96.80%和98.85%。CEX检测结果显示,预洗脱杂质、酸性异构体和原液的主峰含量分别为2.60%、16.77%和66.46%,酸性峰含量分别为97.40%、82.53%和25.92%。c IEF检测结果显示,预洗脱杂质、酸性异构体和原液的主峰含量分别为3.21%、18.17%和33.20%,酸性峰含量分别为94.21%、80.18%和64.38%,p I范围分别为6.40⁸.85、7.698.85、7.69⁸.69和7.948.69和7.94⁸.69。生物活性测定结果显示,预洗脱杂质、酸性异构体和原液的比活性分别为0.45×108.69。生物活性测定结果显示,预洗脱杂质、酸性异构体和原液的比活性分别为0.45×10⁴、1.02×104、1.02×10⁴和0.94×104和0.94×10^{(4 )}U/mg。结论 HER2单克隆抗体酸性组分大部分由于单克隆抗体片段化造成;酸性电荷异构体的存在对HER2单克隆抗体的活性影响并不显著。

关键词(KeyWords)： 人类表皮生长因子受体2;单克隆抗体;酸性电荷异构体

Abstract:

Keywords:

基金项目(Foundation):

作者(Authors): 郭秋菊;李雪;封纯芳;李乐;陈红霞;于玉根;

DOI: 10.13200/j.cnki.cjb.002051

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