在料批发发酵过程中,在化学定义的介质中细胞质生产Fabs
Angel Castillo-Corujo1, Mirva J Saaranen1, Lloyd W Ruddock1
1Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland.
Protein expression and purification
|November 18, 2023
概括
这项研究表明,使用CyDisCo系统在大肠杆菌细胞质中产生高水平的生物活性抗体片段 (Fab) 蛋白质. 这种方法为在工业发酵条件下在微生物系统中生产Fabs提供了可行的替代方案.
科学领域:
- 生物技术是生物技术.
- 蛋白质工程是指蛋白质工程.
- 微生物发酵 微生物发酵
背景情况:
- 抗体片段 (Fab) 蛋白质是具有多种生物医学应用的关键生物分子.
- 哺乳动物细胞表达是全长抗体的标准,但由于它们的结构更简单,Fabs的微生物生产是可行的.
- 对于Fabs而言,大肠杆菌中的周周体表达是常见的,但在二硫化物键形成方面面临挑战.
研究的目的:
- 调查CyDisCo系统对E. coli中Fabs细胞质表达的疗效.
- 在工业相关的发酵条件下实现高水平的生物活性Fabs生产.
主要方法:
- 利用CyDisCo系统在E. coli中形成细胞质二硫化物键.
- 在化学定义的介质中使用料批发发酵来表达两个特定的Fabs (Herceptin和Maa48).
- 分析了蛋白质表达,溶解度,通过CD光谱折叠和生物活性.
主要成果:
- 实现了可溶性Fabs的高产量:Ma48的565 mg/L和Herceptin的660 mg/L.
- 通过循环二元化 (CD) 光谱学证实了适当的蛋白质折叠.
- 表达Fabs.的已证明的生物活性.
结论:
- 赛迪斯科系统使E. coli中生物活性Fabs的有效细胞质生产成为可能.
- 这代表了在工业发酵环境下的微生物系统中高水平的Fab制造的新方法.
- 细胞质表达克服了与Fabs.中围质二硫化物键形成相关的挑战.
相关概念视频
Batch vs Continuous Culture
Fermentation is a foundational biotechnological process used to produce pharmaceuticals, biofuels, enzymes, and food additives. Among industrial strategies, batch and continuous fermentation are the two most widely applied. Although both rely on microbial conversion of substrates into desired products, they differ markedly in operation, productivity, and suitability for specific applications.Batch fermentation occurs in a closed system in which nutrient media and inoculum are added at the...
Fed-Batch Culture
Fed-batch culture is a widely used bioprocessing strategy combining aspects of batch culture with controlled substrate feeding to optimize cell growth and product formation. In this semi-closed system, nutrients are strategically added during fermentation, while the accumulated products and biomass remain within the bioreactor until the end of the operation. This controlled addition of substrates allows for better management of growth kinetics, nutrient limitation, and metabolite...
Upstream Processing
Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
Production of Alcohol
Continuous fermentation is a key strategy in industrial ethanol production, particularly when efficiency, scalability, and high yields are essential. This approach allows for uninterrupted operation and optimized resource utilization. The primary feedstock, corn starch, undergoes enzymatic hydrolysis facilitated by α-amylase and glucoamylase. These enzymes break down the starch into fermentable sugars such as glucose, which are readily assimilated by fermentative microorganisms.Fermentation...
Production of Antibiotics
Penicillin, one of the earliest and most widely used antibiotics, is produced industrially by the filamentous fungus Penicillium chrysogenum. Large stirred-tank bioreactors ranging from tens to hundreds of thousands of liters maintain tightly controlled temperature, pH, and dissolved oxygen conditions to support fungal metabolism and maximize antibiotic yield. Penicillin is a secondary metabolite, synthesized primarily during the stationary growth phase, which requires a carefully managed...


