Solubilization and refolding of variety of inclusion body proteins using a novel formulation

Formation of protein aggregates as inclusion bodies (IBs) still poses a major hurdle in the recovery of bioactive proteins from E. coli. Despite the development of many mild solubilization buffers in last two decades, highthroughput recovery of functional protein from wide range of IBs is still a challenge at an academic and industrial scale. Herein, a novel formulation for improved recovery of bioactive protein from variety of bacterial IBs is developed. This novel formulation is comprised of 20% trifluoroethanol, 20% n-propanol and 2 M urea at pH 12.5 which disrupts the major dominant forces involved in protein aggregation. An extensive comparative study of novel formulation conducted on different IBs demonstrates its high solubilization and refolding efficiency. The overall yield of bioactive protein from human growth hormone expressed as bacterial IBs is reported to be around 50%. This is attributed to the capability of novel formulation to disrupt the tertiary structure of the protein while protecting the secondary structure of the protein, thereby reducing the formation of soluble aggregates during refolding. Thus, the formulation can eliminate the need of screening and optimizing various solubilization formulation and will improve the efficiency of recovering bioactive protein from variety of IB aggregates.

Automated summary

A novel formulation comprised of trifluoroethanol, n-propanol, and urea disrupts protein aggregation forces, enabling high solubilization and refolding efficiency for recovering bioactive proteins from bacterial inclusion bodies. Extensive comparative studies demonstrate a 50% yield of bioactive proteins with the capability to disrupt tertiary protein structures and protect secondary structures, reducing soluble aggregate formation during refolding. This formulation eliminates the need for screening and optimizing solubilization formulations, improving efficiency in recovering bioactive proteins.