在抗体药物产品中检测宿主细胞微蛋白杂质
Ioanna Tzani1, Marina Castro-Rivadeneyra1,2, Paul Kelly1
1National Institute for Bioprocessing Research and Training, Fosters Avenue, Blackrock, Co, Dublin, Ireland.
Nature communications
|October 4, 2024
概括
研究人员在中国仓鼠卵巢 (CHO) 细胞中发现了数千种新型微蛋白,这些细胞对于产生治疗性抗体至关重要. 这些微蛋白可能是抗体药物的杂质,它们的水平在细胞培养过程中发生变化.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 蛋白质组学是指蛋白质组学.
背景情况:
- 中国仓鼠卵巢 (CHO) 细胞是产生治疗性单克隆抗体 (mAbs) 和Fc融合蛋白的主要宿主.
- 在CHO细胞中对非正规翻译事件的不完整注释阻碍了对细胞生物学的理解和对宿主细胞蛋白质 (HCP) 杂质的检测.
研究的目的:
- 在CHO细胞中使用核糖体足迹分析 (Ribo-seq) 识别新型的开放式读取框架 (ORF) 和微蛋白.
- 为了调查商业抗体药物产品中微蛋白质杂质的存在.
- 了解CHO细胞培养中的微蛋白丰度的调节.
主要方法:
- 使用核糖体足迹分析 (Ribo-seq) 发现了新的ORF,包括N端延伸和短ORF (sORF).
- 对商业抗体药物产品进行了基于质谱的HCP分析,使用扩展蛋白序列数据库.
主要成果:
- 在CHO细胞中发现了数千种新型ORF,包括编码微蛋白的sORF.
- 在8种商业抗体药物产品 (7mAbs,1Fc-fusion) 中检测到微蛋白质杂质.
- 发现微蛋白的丰富程度因细胞生长阶段和培养条件而异.
结论:
- 这项研究通过识别众多微蛋白质,显著扩大了已知的CHO细胞蛋白质组.
- 这些发现揭示了微蛋白质是治疗性抗体产品中潜在的HCP杂质.
- 这项研究为研究CHO细胞中非正典翻译和蛋白质合成调节提供了宝贵的资源.
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