通过递归高压同质化增加细菌包容体的纯度和重新折叠产量
Robert Klausser1, Lukas Veiter2, Julian Kopp1
1Research Division Integrated Bioprocess Development, Institute of Chemical, Environmental and Bioscience, Technische Universität Wien, Vienna, Austria; Christian Doppler Laboratory IB Processing 4.0, Technische Universität Wien, Vienna, Austria.
Journal of biotechnology
|November 7, 2025
概括
一种新的递归高压均质化方法改善了用于治疗性蛋白质重新折叠的包容体纯度. 这种方法提高了产品的产量和可持续性,减少了对环境的影响.
科学领域:
- 生物技术是生物技术.
- 生物工艺工程 生物工艺工程
- 蛋白质治疗药物 蛋白质治疗药物
背景情况:
- 治疗性蛋白质通常以包括体的形式在大肠杆菌中产生.
- 目前使用批量稀释的重新折叠方法是低效和不可持续的,因为蛋白质度低,用水量高,设备要求大.
研究的目的:
- 开发和评估一种"递归高压均质化"的方法,以优化细胞溶解和纳入体洗.
- 在溶解和重新折叠之前,提高纳入体的纯度.
- 增加最终的重新折叠蛋白质度,提高工艺的可持续性.
主要方法:
- 在七种工艺变化中比较了"递归高压均化"与传统的"线性冲洗".
- 评估了细胞溶解,核酸释放和纳入体的纯度.
- 溶解并重新折叠的纳入体,以确定下游加工对产品度的影响.
主要成果:
- 循环高压均质化方案在重新折叠后实现了最高产品度855 mg/L.
- 这种方法使得尿素的二氧化碳排放量减少了约18%.
- 与线性洗相比,每种生物质的产品产量从5.17μg/kg增加到7.84μg/kg.
结论:
- 递归高压均质化是一种有效的策略,可以提高包含体的纯度,并提高治疗性蛋白质重新折叠的产量.
- 开发的方法提供了一个更可持续,更有效的生物处理方法,不需要专门的设备或化学品.
- 优化早期下游加工步骤对最终产品度和环境足迹产生重大影响.
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