School of Interdisciplinary Research, Indian Institute of Technology Delhi, New Delhi, India.
Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi, India.
Methods Mol Biol. 2023;2617:249-256. doi: 10.1007/978-1-0716-2930-7_18.
Escherichia coli is an industrial-relevant microbial host system, which is highly preferred for the large-scale production of recombinant biotherapeutics. Overexpression of these recombinant biotherapeutics in the E. coli system often results in the formation of insoluble protein aggregates termed as inclusion bodies (IBs). The yield and quality of IBs are affected by a spectrum of parameters like temperature, optical density, medium composition, induction time, and amount of inducer. Here, we present a protocol for the formation and processing of IBs for production of recombinant human granulocytes colony-stimulating factor (rGCSF).
大肠杆菌是一种与工业相关的微生物宿主系统,非常适合大规模生产重组生物疗法。在大肠杆菌系统中过度表达这些重组生物疗法通常会导致形成不溶性蛋白质聚集体,称为包涵体 (IBs)。IBs 的产量和质量受一系列参数的影响,如温度、光密度、培养基成分、诱导时间和诱导剂用量。在这里,我们提供了一个用于形成和处理 IBs 以生产重组人粒细胞集落刺激因子 (rGCSF) 的方案。