Optimization of Refolding of Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor (rhGM-CSF) Immobilized on Affinity Sorbent

Автор: 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, 1605 просмотров.