Expression of recombinant human mutant granulocyte colony stimulating factor (Nartograstim) in Escherichia coli.
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.
Centro de Biotecnologia, Instituto Butantan, Av Vital Brasil 1500, São Paulo, SP, 05503-900, Brazil.
F R
FR
Gomes
A C
AC
Maluenda
J O
JO
Tápias
F L S
FL
Oliveira
L C
LC
Sá-Rocha
E
E
Carvalho
P L
PL
Ho
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor