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Optimization of Refolding of Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor (rhGM-CSF) Immobilized on Affinity Sorbent
Optimization of Refolding of Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor (rhGM-CSF) Immobilized on Affinity SorbentАвтор: E.N. Kosobokova, K.A. Skrypnik, M.V. Pinugina, A.I. Shcherbakov, and V.S. Kosorukov Страница: 39-46
Optimization of Refolding of Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor (rhGM-CSF) Immobilized on Affinity Sorbent Biotekhnologiya, 2013, N 3, P. 39-46 UDC 57.088.2 Section: “Biologicals Technology”
E.N. Kosobokova *, K.A. Skrypnik, M.V. Pinugina, A.I. Shcherbakov, and V.S. Kosorukov The Blokhin Russian Oncological Research Center, Russ. Acad. Med. Sci., 115478, Moscow Russia e-mail: ekkos@mail.ru
A method for obtaining and purification of biologically active recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) synthesized as a result of the expression in Escherichia coli cells has been developed. A pre-washing of inclusion bodies fraction which allows to remove numerous ballast proteins, and on-column refolding of rhGM-CSF are specific characteristics of the method. The refolding effeciency reached 70%, wereas the target preparation purity was 95%. The biological activity of the purified and refolded protein was similar to those of other recombinant hGM-CSF analogues.
Key words: GM-CSF, His-tag, on-column refolding.
The full English version of the article was published in “Applied Biochemistry and Microbiology”, 2014, Vol. 50, Issue 8, pp. 773-779 as E. N. Kosobokova, K. A. Skrypnik, M. V. Pinyugina, A. I. Shcherbakov, and V. S. Kosorukov “Optimization of the refolding of recombinant human granulocyte-macrophage colony-stimulating factor immobilized on affinity sorbent”. It is contained at the SpringerLink website: http://link.springer.com/article/10.1134/S0003683814080031 DOI: 10.1134/S0003683814080031
09.04.2015, 1618 просмотров. |
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