Renaturation of recombinant human neurotrophin‐3 from inclusion bodies using a suppressor agent of aggregation

Abstract

Escherichia coli has been widely used in the production of recombinant proteins. One of the drawbacks inherent in this method is that the proteins produced in the cells often form inactive inclusion bodies. Usually, the inclusion bodies can be separated from other cell components, solubilized by denaturants such as guanidine hydrochloride or urea, and then renatured through a refolding process such as dilution or dialysis. However, it has been shown that biologically active recombinant human neurotrophin‐3 cannot be obtained at high yield by this procedure due to aggregation and precipitation of the protein. We applied the refolding process using the aggregation suppressor L‐arginine in the renaturation of neurotrophin‐3, and obtained biologically active neurotrophin‐3 at high yield from the inclusion bodies. Consequently, about 10 mg of purified neurotrophin‐3 was prepared from 1 litre of culture broth.