Atousa Aliahmadi - Biotechnology Dept,.Pasteur Institute of Iran, Tehran, Iran
ali Moradi,
Maryam Omidi
Soroush Sardari

In this study a rapid, single – step purification method for recombinant human proinsulin, expressed in E. coli cells is reported and the subsequent refolding procedure and characterization of the refolded product are described. The inclusion bodies (IBs) of protein A fusion human proinsulin were solubilized and converted to hexasulfonate derivatives. Then the fusion protein was purified using a modified acid precipitation protocol up to 80 % purity. Purified fusion protein was refolded by two methods and the efficiency of these methods was evaluated using HPLC and native PAGE. The best reaction time for refolding step was determined to be about 16 hours.

Recombinant proinsulin, Purification, Refolding, E. coli