Zahra Hajihassan, Mehri Abdi, Elaheh Roshani Yasaghi, Azra Rabbani-Chadegani
||Minerva Biotecnologica, 2017, Vol.29 (3)
BACKGROUND: Several proteins have been purified in a single step from E. coli using fused histidine tails and immobilized metal affinity chromatography (IMAC). Many attempts have been done in order to optimize protein purification by this method. METHODS: As the efficiency of purification is dependent on buffers and conditions used, so in this study different pH, imidazole concentration and incubation time were surveyed to optimize recombinant Î²-NGF purification. RESULTS: In this work pET39b vector containing DsbA sequence was used in order to produce recombinant Î²-NGF with his tag tails; Î²-NGF is produced in the form of DsbA-Î²NGF fusion by using this vector. Also in order to determine the secondary structure and function of purified Î²-NGF, CD spectroscopy and treatment of PC12 cell... line with Î²-NGF was done. CONCLUSIONS: Our data represented the best strategies to increase the purification efficiency using IMAC. PC12 treatment with both purified recombinant Î²-NGF and DsbA-Î²NGF fusion indicated differentiation of this cell line to the nerve cells, demonstrating that DsbA-Î²NGF fusion retained Î²-NGF natural activity and is fully functional.