替代拼接用于可调节的蛋白质子单元的表达在所需的比率
Christel Aebischer-Gumy1, Pierre Moretti1, Timothee Brunstein Laplace1
1Drug Substance Development, Ichnos Sciences SA (formerly Glenmark Pharmaceuticals SA), La Chaux-de-Fonds, Switzerland.
mAbs
|April 23, 2024
概括
本研究介绍了一种替代拼接方法,用于控制单基因的蛋白质比率. 这种方法使得可调节的表达能够有效地产生多重蛋白质,包括双特异性抗体.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 蛋白质工程是指蛋白质工程.
背景情况:
- 在稳定的比例下控制多种蛋白质的表达具有挑战性,特别是对于像双特异性抗体这样的多重蛋白质.
- 低于最佳的子单位比率阻碍了高效的组装,并增加了副产品的形成.
研究的目的:
- 开发一种新的方法,使用替代拼接来调整和预定义来自单个促进子的两个不同的多链的表达比.
- 为了证明这种方法用于生产光蛋白和复杂的多重蛋白质,包括双特异性抗体的应用.
主要方法:
- 采用了含有两个前子和侧面类热素4 (cTNT-I4) 序列的前mRNA的替代拼接.
- 工程拼接接受器强度调节拼接聚链的比率.
- 光蛋白 (GFP, dsRED) 和双特异抗体子单元的构造表达载体.
- 使用滴滴数字PCR进行拼接比率稳定性评估和完整质量分析以评估产品质量.
主要成果:
- 在14天的食批次过程中,实现了多重蛋白质的工业相关表达水平 (>1 g/L).
- 在28天的培养期内表现出稳定的拼接比率.
- 已确认的产品质量,没有不必要的接相关杂质.
- 成功表达了一种BEAT®类型双特异抗体的子单位.
结论:
- 替代拼接提供了一种强大的工具,可以精确控制蛋白质子单元固体测量.
- 这种方法促进了复杂的多重蛋白质的高效生产,如双特异性抗体,具有高质量和产量.
- 开发的系统具有多功能性,适用于生物技术中的各种蛋白质表达挑战.
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