%0 Journal Article %A Mahmoodi, Merat %A Ghodsi, Maryam %A Moghadam, Malihe %A Sankian, Mojtaba %T Pulsed Dilution Method for the Recovery of Aggregated Mouse TNF-α %J Reports of Biochemistry and Molecular Biology %V 5 %N 2 %U http://rbmb.net/article-1-109-en.html %R %D 2017 %K Escherichia coli, Guanidine Hydrochloride, Inclusion Bodies, Mouse TNF-(α), %X Background: The expression of mouse tumor necrosis factor alpha (TNF-α) in Escherichia coli is a favorable way to get high yield of protein; however, the formation of cytoplasmic inclusion bodies, which is the consequence of insoluble accumulated proteins, is a major obstacle in this system. To overcome this obstacle, we used a pulsed dilution method to convert the product to its native conformation. Methods: Reducing agent and guanidine hydrochloride were used to solubilize inclusion bodies formed after TNF-(α) expression. Then, the refolding procedure was performed by pulsed dilution of the denatured protein into a refolding buffer. The properly-folded protein was purified by metal affinity chromatography. Results: SDS-PAGE showed a 19.9 kDa band related to the mature TNF-(α) protein. The protein was recognized by anti-mouse TNF-(α) on western blots. The final concentration of the purified recombinant TNF-(α) was 62.5 µg/mL. Conclusions: Our study demonstrates the efficiency of this method to produce a high yield of folded mature TNF- (α). %> http://rbmb.net/article-1-109-en.pdf %P 103-107 %& 103 %! %9 Original Article %L A-10-1-73 %+ Immuno-Biochemistry lab, Immunology Research Center, Buali Research Institute , School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. %G eng %@ 2322-3480 %[ 2017