Abstract
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.
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Original Russian Text © T.V. Fedorov, V.I. Korobov, V.G. Nazarov, A.E. Smolkina, V.A. Shmelev, 2010, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2010, Vol. 46, No. 2, pp. 243–247.
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Fedorov, T.V., Korobov, V.I., Nazarov, V.G. et al. Purification of recombinant proteins with an example of tumor necrosis factor thymosin-α1. Appl Biochem Microbiol 46, 226–229 (2010). https://doi.org/10.1134/S0003683810020171
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DOI: https://doi.org/10.1134/S0003683810020171