• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biotechnology Progress >Evaluation of a Novel Methacrylate-Based Protein A Resin for the Purification of Immunoglobulins and Fc-Fusion Proteins
【24h】

Evaluation of a Novel Methacrylate-Based Protein A Resin for the Purification of Immunoglobulins and Fc-Fusion Proteins

机译：新型基于甲基丙烯酸酯的蛋白A树脂对免疫球蛋白和Fc融合蛋白的纯化的评估

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

Protein A affinity chromatography is a central part of most commercial monoclonal antibody and Fc-fusion protein purification processes. In the last couple years an increasing number of new Protein A technologies have emerged. One of these new Protein A technologies consists of a novel, alkaline-tolerant, Protein A ligand coupled to a macroporous polymetha-crylate base matrix that has been optimized for immunoglobulin (Ig) G capture. The resin is interesting from a technology perspective because, the particle size and pore distribution of the base beads are reported to have been optimized for high IgG binding and fast mass transfer, while the Protein A ligand has been engineered for enhanced alkaline tolerance. This resin was subjected to a number of technical studies including evaluating dynamic and static binding capacities, alkaline stability, Protein A leachate propensity, impurity clearance, and pressure—flow behavior. The results demonstrated similar static binding capacities as those achieved with industry standard agarose Protein A resins, but marginally lower dynamic binding capacities. Removal of impurities from the process stream, particularly host cell proteins, was molecule dependent, but in most instances matched the performance of the agarose resins. This resin was stable in 0.1 M NaOH for at least 100 h with little loss in binding capacity, with Protein A ligand leakage levels comparable to values for the agarose resins. Pressure-flow experiments in lab-scale chromatography columns demonstrated minimal resin compression at typical manufacturing flow rates. Prediction of resin compression in manufacturing scale columns did not suggest any pressure limitations upon scale up.
机译：蛋白A亲和色谱是大多数商业单克隆抗体和Fc融合蛋白纯化工艺的核心部分。在最近几年中，出现了越来越多的新蛋白质A技术。这些新的Protein A技术之一是由一种新颖的，耐碱性的Protein A配体与大孔聚甲基丙烯酸甲酯基础基质偶联而成，该基质已针对免疫球蛋白（Ig）G捕获进行了优化。从技术角度来看，该树脂很有趣，因为据报道，基础珠的粒径和孔分布已针对高IgG结合和快速传质进行了优化，而蛋白A配体已针对增强的碱耐受性进行了设计。对该树脂进行了许多技术研究，包括评估动态和静态结合能力，碱性稳定性，蛋白A浸出液的倾向性，杂质清除率以及压力-流动行为。结果表明，静态结合能力与工业标准琼脂糖蛋白A树脂所获得的类似，但动态结合能力稍低。从工艺流中去除杂质，尤其是宿主细胞蛋白，是依赖于分子的，但是在大多数情况下，与琼脂糖树脂的性能相匹配。该树脂在0.1 M NaOH中稳定至少100小时，结合能力几乎没有损失，蛋白质A配体的泄漏水平与琼脂糖树脂的值相当。在实验室规模的色谱柱上进行的压力流实验表明，在典型的制造流速下，树脂压缩最小。在生产规模的色谱柱中对树脂压缩的预测并未表明在规模放大时存在任何压力限制。

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