...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Biotechnology >Analysis of changes during subclone development and ageing of humanantibody-producing heterohybridoma cells by Northern blot and flowcytometry
【24h】

Analysis of changes during subclone development and ageing of humanantibody-producing heterohybridoma cells by Northern blot and flowcytometry

机译：通过Northern印迹和流式细胞仪分析产生人抗体的异种杂交瘤细胞亚克隆发育和衰老过程中的变化

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The economic importance of obtaining high-producing subclones for large scale production of pharmaceutical proteins is self-evident. However, few papers have studied the changes that occur during subclone development. This information would be important for further improvement of screening and subcloning protocols. We have therefore compared subclones of a human-mouse heterohybridoma cell line producing a human antibody against HIV-1. Three subclones with low, medium and high specific production rates were selected for this study and their light and heavy chain mRNA content, the intracellular content of light and heavy chain and the specific secretion rates compared. In addition the long time stability of antibody expression in the highest producing subclone was analysed for one year. For the three subclones a correlation between the intracellular content in light chain and the secretion rate was found, while the intracellular content in heavy chain was the same for all three subclones. These results indicate that the assembly in the endoplasmic reticulum (ER) is one of the major rate limiting factors in antibody production. During long time cultivation of the heterohybridoma cell line a continuous decrease in light and heavy chain production was seen without the appearance of a non producing sub-population.
机译：不言而喻，获得用于大规模生产药物蛋白的高产量亚克隆的经济重要性。但是，很少有论文研究亚克隆开发过程中发生的变化。此信息对于进一步改进筛选和亚克隆方案非常重要。因此，我们比较了产生抗HIV-1人抗体的人-小鼠异种杂交瘤细胞系的亚克隆。本研究选择了三种具有低，中，高比生产率的亚克隆，并比较了它们的轻链和重链mRNA含量，轻链和重链的细胞内含量以及比分泌率。另外，分析了在最高产量的亚克隆中抗体表达的长期稳定性一年。对于这三个亚克隆，发现轻链中的细胞内含量与分泌速率之间具有相关性，而对于所有三个亚克隆，重链中的细胞内含量均相同。这些结果表明内质网（ER）中的组装是抗体生产中的主要限速因素之一。在异源杂交瘤细胞系的长时间培养过程中，观察到轻链和重链产量的持续下降，而没有出现非生产性亚群。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号