生物仿真亲和色谱用于抗体净化:宿主细胞蛋白质结合和杂质去除
Haotian Huang1, Xiaoyan Dong2, Yan Sun2
1Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China.
Journal of chromatography. A
|August 22, 2023
概括
这项研究开发了一种用于单克隆抗体净化的基亲和色谱法,通过临床应用的综合方法改善宿主细胞蛋白质和DNA去除.
科学领域:
- 生物技术是生物技术.
- 生物分离 生物分离
- 蛋白质化学 蛋白质化学
背景情况:
- 基亲和色谱是一种新兴的替代蛋白A用于抗体净化.
- 较低的选择性,特别是宿主细胞蛋白 (HCP) 和DNA的选择性,限制了其应用.
- 有效的污染物去除对于单克隆抗体 (mAb) 制造至关重要.
研究的目的:
- 合成和评估用于mAb捕获和HCP结合机制的系凝 (SepFF-bPL).
- 为了应对HCP和DNA去除在亲和染色体学中的挑战.
- 为临床级的mAbs.开发一个有效的净化列车.
主要方法:
- 一个生物仿真联体 (bPL) FYWHCLDE与Sepharose 6快速流 (SepFF) 的合.
- 研究SepFF-bPL列上的HCP结合机制,包括静电相互作用和离子导电性的作用.
- 实施一个净化列车,结合色素提取,离子交换吸附和SepFF-bPL色谱.
主要成果:
- 在SepFF-bPL列显示有效的mAb捕获和聚合物去除.
- 最初观察到HCP和DNA的去除不良.
- 机械学研究显示,静电相互作用主导HCP结合,受离子导电性的影响.
- 优化的净化列车实现了高mAb回收 (68.7%),低HCP (23ppm) 和无法检测到的DNA.
结论:
- 基亲和色谱需要优化,以有效地去除污染物.
- 了解HCP绑定机制是制定有效净化策略的关键.
- 整合亲和色谱的多步净化列车可以满足mAb纯度的临床要求.
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