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Expression of recombinant human mutant granulocyte colony stimulating factor (Nartograstim) in Escherichia coli

Author:
Gomes, F. R., Maluenda, A. C., Tápias, J. O., Oliveira, F. L. S., Sá-Rocha, L. C., Carvalho, E., Ho, P. L.
Source:
World journal of microbiology & biotechnology 2012 v.28 no.7 pp. 2593-2600
ISSN:
0959-3993
Subject:
Escherichia coli, bioactive properties, cell proliferation, colony-stimulating factors, humans, markets, mice, polyclonal antibodies
Abstract:
The human granulocyte colony stimulating factor (hG-CSF) plays an important role in hematopoietic cell proliferation/differentiation and has been widely used as a therapeutic agent for treating neutropenias. Nartograstim is a commercial G-CSF that presents amino acid changes in specific positions when compared to the wild-type form, which potentially increase its activity and stability. The aim of this work was to develop an expression system in Escherichia coli that leads to the production of large amounts of a recombinant hG-CSF (rhG-CSF) biosimilar to Nartograstim. The nucleotide sequence of hg-csf was codon-optimized for expression in E. coli. As a result, high yields of the recombinant protein were obtained with adequate purity, structural integrity and biological activity. This protein has also been successfully used for the production of specific polyclonal antibodies in mice, which could be used in the control of the expression and purification in an industrial production process of this recombinant protein. These results will allow the planning of large-scale production of this mutant version of hG-CSF (Nartograstim), as a potential new biosimilar in the market.
Agid:
481180