Increasing the solubility of single-chain variable fragment (scFv) antibody produced in recombinant Escherichia coli by fed-batch culture with DO-stat.

摘要

Protein expression system using E. coli is still a first choice for recombinant protein production (Tripathi 2016). However, proteins expressed in E. coli are frequently misfolded into insoluble forms, so called inclusion bodies. The insoluble proteins require a complicated process for purification including a refolding step, therefore, the increase of solubility of expressed proteins has been a critical issue in recombinant protein production using E. coli. Though there are many strategies concerning the solubility improvement, it is known that the solubility is increased by lowering the protein synthesis rate under proper DO environment. In this report, we challenge increasing the solubility of single-chain variable fragment (scFv) produced in E. coli by applying a fed-batch culture with DO-stat, which allows to control protein synthesis rate under precisely controlled DO environment.