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;2617:201-208. doi: 10.1007/978-1-0716-2930-7_14.
Microbial-based biotherapeutics that are produced in Escherichia coli (E. coli) can be generated intracellularly in the form of inclusion bodies (IBs) or in soluble active form in periplasmic space or extracellularly. Overexpression of these biotherapeutics in E. coli leads to formation of insoluble aggregates called inclusion bodies. These IBs contain misfolded and inactive form of proteins which need to be refolded to obtain a functionally active form of proteins. Here, we discuss refolding of E. coli-based recombinant human granulocyte colony-stimulating factor (GCSF), expressed as IBs, and highlight some of the key features associated with the refolding kinetic reaction.
微生物来源的生物治疗药物在大肠杆菌 (Escherichia coli, E. coli) 中产生时，可以以包涵体 (inclusion bodies, IBs) 的形式在细胞内、周质空间或细胞外以可溶性活性形式产生。这些生物治疗药物在大肠杆菌中的过表达会导致形成不溶性聚集体，称为包涵体。这些包涵体包含错误折叠和无活性的蛋白质，需要重折叠以获得具有功能活性的蛋白质形式。在这里，我们讨论了作为包涵体表达的大肠杆菌来源的重组人粒细胞集落刺激因子 (recombinant human granulocyte colony-stimulating factor, rhGCSF) 的重折叠，并强调了与重折叠动力学反应相关的一些关键特征。
