| 重组单抗药物的非还原电泳纯度分析 |
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| 引用本文： | 程立均,魏敬双,张世雄,李耿,贾茜.重组单抗药物的非还原电泳纯度分析[J].粉末涂料与涂装,2010,23(1). |
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| 作者姓名： | 程立均 魏敬双 张世雄 李耿 贾茜 |
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| 作者单位： | 华北制药集团新药研究开发有限责任公司生物技术研究室,石家庄,050015 |
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| 基金项目： | 863计划生物和医药技术领域重大项目 |
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| 摘 要： | 目的对重组单抗药物的非还原电泳纯度进行分析,排除样品制备过程中形成的一些假象。方法结合重组单抗的分子结构特征,改变常规SDS-PAGE条件,将抗体样品在不同的缓冲液及电泳条件下进行比较。结果抗体在非还原SDS-PAGE纯度检测中出现的次带,可通过在供试品缓冲液中加入烷基化试剂封闭自由巯基而几乎全部消除;通过降低电泳过程的环境温度,可有效降低主带上方高相对分子质量带的出现。结论单抗药物中由于含有多对二硫键引起的一些因电泳样品制备而形成的假象带,可以通过试验方法的修正而去除。
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| 关 键 词： | 重组单抗 非还原SDS-PAGE 纯度 二硫键 |
Analysis of Non-reduced SDS-PAGE Purity of Recombinant Monoclonal Antibody |
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| CHENG Li-jun,WEI Jing-shuang,ZHANG Shi-xiong,et al.Analysis of Non-reduced SDS-PAGE Purity of Recombinant Monoclonal Antibody[J].Chinese Journal of Biologicals,2010,23(1). |
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| Authors: | CHENG Li-jun WEI Jing-shuang ZHANG Shi-xiong |
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| Affiliation: | CHENG Li-jun,WEI Jing-shuang,ZHANG Shi-xiong,et al (New Drug R&D Center,North China Pharmaceutical Corporation,Shijiazhuang 050015,China) |
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| Abstract: | Objective To analyze the non-reduced SDS-PAGE purity of recombinant monoclonal antibody(McAb)and exclude the artifacts formed during preparation of test samples.Methods The condition for routine SDS-PAGE was modified based on the structural characters of recombinant McAb molecules.The recombinant McAb samples were treated with various loading buffers under various conditions for SDS-PAGE,and the results were compared.Results Almost all the minor bands appeared during determination of purity of McAb by non-r... |
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| Keywords: | Recombinant monoclonal antibody Non-reduced SDS-PAGE Purity Disulfide bond |
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