Expressão do fator estimulador de colônia de granulócito humano recombinante (rhG-CSF) em Escherichia coli. / Expression of recombinant human colony stimulating factor (rhG-CSF) in Escherichia coli.

AUTOR(ES)
DATA DE PUBLICAÇÃO

2010

RESUMO

The recombinant human granulocyte colony stimulating factor (rhG-CSF) is a non-glycosylated protein with 175 amino acids. This factor plays an important role in hematopoietic cell proliferation and has been widely used for treating neutropenia. The purpose of this work is to construct two expression systems in E. coli; a system for obtaining rhG-CSF in the cytoplasm and the other for secretion of recombinant protein in the culture medium using the signal sequence of L-asparaginase II. The two expression systems were tested and compared. From these data, the next steps for obtaining the rhG-CSF were done with the expression system without the signal sequence. The refolding and purification steps were efficient, resulting in a protein with adequate purity, structural integrity and biological activity. This protein has also been successfully used for the production of polyclonal antibodies in mice. With these results, the protein rhG-CSF was feasible for further studies in bioreactors and pilot scale production.

ASSUNTO(S)

bacterial genetics escherichia coli genética bacteriana granulocytes cytoplasmic expressão heteróloga heterolog expression rhg-csf rhg-csf fatores biológicos granulocytes (human) granulócitos citoplasmáticos escherichia coli granulócitos (humano) biological factors corpos de inclusão inclusion body

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