Moscow region, Russia
Moscow region, Russia

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Fedorov T.V.,OOO NPP Pharmaklon | Korobov V.I.,OOO NPP Pharmaklon | Nazarov V.G.,OOO NPP Pharmaklon | Smolkina A.E.,OOO NPP Pharmaklon | Shmelev V.A.,OOO NPP Pharmaklon
Applied Biochemistry and Microbiology | Year: 2010

Hybrid protein, cancer necrosis factor thymosin-α1 (TNF-T), when synthesizing in strain-producer of Escherichia coli SG200-50 with plasmid pThy315, was a part of "inclusion bodies" mostly in the form of a high-molecular complex with other proteins due to the S-S bonds formation. An approach of purification of TNF-T has been proposed, which is based on the destruction of the complex in the presence of sodium dodecylsulfate (DDS-Na) and dithiotreitol (DDT) followed by gel-filtration on Sephadex G-100 and renaturation by ultrafiltration on hollow fibers. The method allows the isolation of electrophoretically homogeneous TNF-T containing no DDS-Na and having high cytotoxic activity against cancer cells of mouse adenocarcinome L-929. The yield of TNF-T achieved 80% relative its content in biomass and 30% relative the total protein. © 2010 Pleiades Publishing, Ltd.

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