title: Recombinant glucagon: a differential biological activity.
abstract: In Brazil, there is a growing demand for specialised pharmaceuticals, and the high cost of their importation results in increasing costs, reaching US$ 1.34 billion in 2012 and US$ 1.61 billion in 2013. Worldwide expenses related to drugs could reach US$ 1.3 trillion in 2018, especially due to new treatments for hepatitis C and cancer. Specialised or high-cost pharmaceutical drugs used for the treatment of viral hepatitis, multiple sclerosis, HIV and diabetes are distributed free of charge by the Brazilian government. The glucagon peptide was included in this group of high-cost biopharmaceuticals in 2008. Although its main application is the treatment of hypoglycaemia in diabetic patients, it can also be used with patients in an alcoholic coma, for those patients with biliary tract pain, and as a bronchodilator. Therefore, in order to reduce biopharmaceutical production costs, the Brazilian government passed laws focusing on the development and increase of a National Pharmaceutical Industrial Centre, including the demand for the national production of glucagon. For that reason and given the importance and high cost of recombinant glucagon, the purpose of this study was to develop methods to improve production, purification and performance of the biological activity of recombinant glucagon. Glucagon was recombined into a plasmid vector containing a Glutathione S-transferase tag, and the peptide was expressed in a heterologous Escherichia coli system. After purification procedures and molecular analyses, the biological activity of this recombinant glucagon was examined using in vivo assays and showed a highly significant (p < 0.00001) and prolonged effect on glucose levels when compared with the standard glucagon. The experimental procedure described here facilitates the high level production of recombinant glucagon with an extended biological activity.
authors: Department of Molecular Pathology, University of Brasilia, Brasilia, DF Brazil ; Laboratory of Plant-Pest Interaction, Embrapa - Genetic Resources and Biotechnology, Brasília, DF Brazil.
authors: Angelina M M
authors: AM
authors: Basso
authors: Laboratory of Plant-Pest Interaction, Embrapa - Genetic Resources and Biotechnology, Brasília, DF Brazil.
authors: Patrícia B
authors: PB
authors: Pelegrini
authors: Laboratory of Plant-Pest Interaction, Embrapa - Genetic Resources and Biotechnology, Brasília, DF Brazil ; Pioneer Union for Social Insertion - UPIS, Planaltina, DF Brazil.
authors: Fernanda
authors: F
authors: Mulinari
authors: Laboratory of Plant-Pest Interaction, Embrapa - Genetic Resources and Biotechnology, Brasília, DF Brazil.
authors: Michelle C
authors: MC
authors: Costa
authors: Laboratory of Plant-Pest Interaction, Embrapa - Genetic Resources and Biotechnology, Brasília, DF Brazil ; Catholic University of Brasilia, Brasilia, DF Brazil.
authors: Antonio B
authors: AB
authors: Viana
authors: Laboratory of Mass Spectrometry, Embrapa - Genetic Resources and Biotechnology, Brasília, DF Brazil.
authors: Luciano P
authors: LP
authors: Silva
authors: Laboratory of Plant-Pest Interaction, Embrapa - Genetic Resources and Biotechnology, Brasília, DF Brazil ; Catholic University of Brasilia, Brasilia, DF Brazil.
authors: Maria Fatima
authors: MF
authors: Grossi-de-Sa
keywords: Biological activity; Enterokinase; GST tag; Heterologous system; Peptide expression; Recombinant Glucagon