色谱 ›› 2019, Vol. 37 ›› Issue (6): 666-670.DOI: 10.3724/SP.J.1123.2018.12023

• 技术与应用 • 上一篇    

十二烷基硫酸钠毛细管电泳法分析非还原单克隆抗体药物纯度的前处理条件优化

刘振东2, 高铁1, 徐玲丽1, 杨勇2, 王庆民2, 陈泓序1   

  1. 1. CE & Biopharma, Sciex, 北京 100015;
    2. 齐鲁制药有限公司药物研究院, 山东 济南 250100
  • 收稿日期:2018-12-25 出版日期:2019-06-08 发布日期:2015-04-17
  • 通讯作者: 王庆民,E-mail:hongxu.chen@sciex.com;陈泓序,E-mail:qingmin.wang@qilu-pharma.com.

Optimization of sample preparation condition for sodium dodecyl sulfate capillary electrophoresis of non-reduced monoclonal antibodies

LIU Zhendong2, GAO Tie1, XU Lingli1, YANG Yong2, WANG Qingmin2, CHEN Hongxu1   

  1. 1. CE & Biopharma, Sciex, Beijing 100015, China;
    2. Drug Research Institute of Qilu Pharmaceutical Co., Ltd., Jinan 250100, China
  • Received:2018-12-25 Online:2019-06-08 Published:2015-04-17

摘要:

十二烷基硫酸钠毛细管电泳法因具有快速、分辨率高的优势,已成为单克隆抗体纯度分析的主流方法。在非还原单克隆抗体纯度检测中,其样品前处理过程对于结果有显著影响。为优化样品前处理,考察了以碘乙酰胺和N-乙基马来酰亚胺为巯基封闭剂,在pH 6.0~9.0的样品缓冲液条件下,不同种类和批次的单抗的纯度。发现在两种巯基封闭剂中,高pH的样品缓冲液会影响巯基封闭的效果,产生较多的抗体片段;而在低pH条件下,抗体片段较少,单抗的纯度更高。因此在进行非还原单抗的纯度检测中,pH 6.0的样品缓冲液为最佳的前处理条件。

关键词: N-乙基马来酰亚胺, 单克隆抗体, 碘乙酰胺, 非还原, 毛细管电泳法, 十二烷基硫酸钠

Abstract:

Sodium dodecyl sulfate capillary electrophoresis has become the mainstream method for purity analysis of monoclonal antibodies because of its advantages of fast and high resolution. Sample preparation has a significant impact on the purity detection of non-reduced monoclonal antibodies. In order to optimize sample preparation, the purity of monoclonal antibodies of different types and batches in sample buffers with iodoacetamide and N-ethyl maleimide as sulfhydryl sealants and at pH 6.0-9.0 was investigated. It was found that in the two types of sulfhydryl sealants, the high pH sample buffer could affect the sealing effect of the sulfhydryl sealant and produce more antibody fragments. Conversely, under the low pH condition, the antibody fragments were fewer and the purities of monoclonal antibodies were higher. Therefore, the sample buffer with pH 6.0 is the optimal preparation condition for the purity detection of non-reduced monoclonal antibodies.

Key words: N-ethylmaleimide (NEM), capillary electrophoresis (CE), iodoacetamide (IAM), monoclonal antibody, non-reduced, sodium dodecyl sulfate (SDS)

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