在基因组最小化Bacillus subtilis菌株中产生抗体
Tobias Schilling1, Rebekka Biedendieck2, Rafael Moran-Torres3
1University Medical Center Groningen, Department of Medical Microbiology, University of Groningen, Hanzeplein 1, P.O. Box 30001, 9700RB Groningen, The Netherlands.
ACS synthetic biology
|February 27, 2025
概括
基因组缩小的细菌菌株,Midi和MiniBacillus,现在可以产生功能性抗体片段. 这一突破克服了蛋白质折叠和分泌抗体的二硫化键形成的先前挑战.
科学领域:
- 生物技术是生物技术.
- 微生物工程 微生物工程
- 蛋白质生产 蛋白质生产
背景情况:
- 细菌细菌是一种成熟的细菌细胞工厂,用于工业酶生产.
- 在B. subtilis中产生复杂的蛋白质,如抗体格式,由于折叠和二硫化键的要求而具有挑战性.
研究的目的:
- 研究使用基因组缩小的Midi和MiniBacillus菌株生产功能抗体碎片 (scFvs,scFabs) 的可行性.
- 为了优化工作流程,以获得高质量的抗体碎片分泌.
主要方法:
- 选择最佳的底盘菌株,表达载体,信号和生长介质.
- 将scFv生产升级到1L生物反应器水平.
- 使用质谱学,生物层干涉学和其他生物物理技术来验证抗体功能.
主要成果:
- MidiBacillus菌株成功分泌了功能性抗体碎片.
- 分泌的抗体碎片表现出本地折叠和正确的二硫化物结合.
- 抗体碎片显示出热稳定性和功能性,如人类C反应蛋白结合scFv.
结论:
- 基因组缩小的Bacillus subtilis菌株适用于高质量的抗体片段分泌.
- 这一进步使得使用微生物系统生产复杂的治疗性蛋白质成为可能.
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