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Methods of preparation of recombinant cytokine proteins
V E Klyushnichenko1, R V Tikhonov, S E Pechenov
1Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklikho-Maklaya 16/10, Moscow, GSP-7, 117871, Russia.
Protein Expression and Purification
|October 29, 1998
Abstract:
Two schemes for efficient and productive isolation for mutant human recombinant tumor necrosis factor-alpha (TNF-alpha R32H) from Escherichia coli cells were developed. The methods include membrane filtration, ion-exchange chromatography and gel filtration, and centrifugation with subsequent free-flow electrophoresis as an alternative procedure. The target product was obtained as active trimer with total yield more than 50% and greater than 98% purity according to PAGE, size-exclusion chromatography, HPLC, and HPCE.