• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Separation and Purification Technology >Partitioning of human antibodies in polyethylene glycol-sodium citrate aqueous two-phase systems
【24h】

Partitioning of human antibodies in polyethylene glycol-sodium citrate aqueous two-phase systems

机译：人抗体在聚乙二醇-柠檬酸钠水两相系统中的分配

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

Partitioning and purification studies of human IgG were performed in polyethylene glycol (PEG) and citrate aqueous two-phase systems (ATPSs). Initial studies performed with pure IgG solutions showed that increasing the concentration of a neutral salt, such as NaCl up to 15% (w/w), increased the partition coefficient of IgG from 0.14 to 5.21. This partitioning behaviour was also observed first in the presence of a simulated protein mixture, containing albumin and myoglobin, and then in a hybridoma cell culture supernatant. Thus, by changing the concentration of NaCl, it is possible to target the partition of IgG to the phase with fewer impurities. The partial purification of IgG from a hybridoma cell culture supernatant in a serum-containing media was achieved using 15% NaCl and recovering the antibody in the top phase. Using an ATPS composed of 8% (w/w) PEG 3350, 8% (w/w) citrate and 15% (w/w) NaCl at pH 6, IgG was recovered with a 99% yield, 44% HPLC purity and with a IgG/protein ratio of 0.9. IgG can be re-extracted to a new citrate phase by decreasing the overall concentration of NaCl to 5%. This back-extraction step introduces not only a further purification step but also allows the separation of the antibody from the polymer. IgG was recovered with a global yield of 99%, with a HPLC purity of 76% and with a IgG/protein ratio of 0.96.
机译：人IgG的分配和纯化研究在聚乙二醇（PEG）和柠檬酸盐水相两相系统（ATPSs）中进行。使用纯IgG溶液进行的初步研究表明，将中性盐（例如NaCl）的浓度提高到15％（w / w），会使IgG的分配系数从0.14增加到5.21。还首先在包含白蛋白和肌红蛋白的模拟蛋白质混合物存在下观察到了这种分配行为，然后在杂交瘤细胞培养上清液中观察到了这种分配行为。因此，通过改变NaCl的浓度，可以将IgG的分配靶向到杂质较少的相。使用15％NaCl从杂交瘤细胞培养上清液中含血清的培养基中部分纯化IgG，并在顶层回收抗体。使用pH为6的，由8％（w / w）PEG 3350、8％（w / w）柠檬酸盐和15％（w / w）NaCl组成的ATPS，以99％的收率，44％HPLC纯度和IgG /蛋白质比为0.9。通过将NaCl的总浓度降低到5％，可以将IgG重新提取到新的柠檬酸盐相中。该反萃取步骤不仅引入了进一步的纯化步骤，而且使抗体与聚合物分离。回收的IgG的总产率为99％，HPLC纯度为76％，IgG /蛋白质比为0.96。

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