Development of Solubilization and Refolding Buffers Publisher Pubmed



Roufarshbaf M¹ ; Akbari V¹ Show Affiliations
Source: Methods in molecular biology (Clifton, N.J.) Published:2023

Abstract

Overexpression of heterologous protein in prokaryotic host cells, such as Escherichia coli, usually leads to formation of inactive and insoluble aggregates known as inclusion bodies (IBs). Recovery of refolded and functionally bioactive proteins from IBs is a challenging task, and a unique condition (e.g., solubilizing and refolding buffers) for each individual protein should be experimentally obtained. Here, we present a simple protocol for development of solubilizing and refolding buffers for successful recovery of pure bioactive proteins from IBs. © 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.