Development of fermentation in a bioreactor for the production of interferon ±-2a secreted into the periplasmic space of Escherichia coli

• Grant number: 15/10928-6
• Support Opportunities: Scholarships in Brazil - Master
• Start date: August 01, 2015
• End date: January 31, 2017
• Field of knowledge: Interdisciplinary Subjects
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Carlos Roberto Jorge Soares
• Grantee: Paulo Victor Sarmento Dias
• Host Institution: Instituto de Pesquisas Energéticas e Nucleares (IPEN). Secretaria de Desenvolvimento Econômico (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

iN 1986, Interferon was the first drug to demonstrate beneficial effects in patients with hepatitis C. Currently, all of hepatitis C treatments are based on the use of combined preparations of IFN-± with or without ribavirin. The therapy results in rapid decline of hepatitis C virus levels in the serum and, in long term, its extinction occurs in blood and liver. The alpha-2 interferon (IFN-±2) are part of a heterologous group of proteins with similar biological activities. The three major IFN-±2 subtypes are ±-2a, ±-2b, ±-2c. The main objective of this project is the production of IFNa-2a in laboratory scale from E. coli bacteria genetically engineered. For this purpose, some intermediate goals need to be achieved. The first of these goals is to validate a method of qualitative and quantitative analysis by reverse phase HPLC for assessing the IFN ±-2a present in bacterial crude extract obtained from osmotic shock. This technique is extremely useful in the next steps, such as the optimization of a fast and efficient fermentation process in a bioreactor for the production of IFN ±-2a using the W3110 strain transformed with the vector LambdaPl-DsbA-IFN±-2a. This strain, developed in our laboratory, is able to secrete interferon in bacterial periplasmic space, so it is in its authentic form. Another goal of this project is to perform the purification of the IFNa-2a in laboratory scale. The final product, after physicochemical and biological characterization may be used as a benchmark for future studies, including potential therapeutic applications. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DIAS, PAULO V. S.; ARTHUSO, FERNANDA S.; OLIVEIRA, JOAO E.; SUZUKI, MIRIAM F.; SOUSA, JOSE M.; RIBELA, MARIA TERESA C. P.; BARTOLINI, PAOLO; SOARES, CARLOS R. J.. Determination of recombinant Interferon-alpha 2 in E. coil periplasmic extracts by reversed-phase high-performance liquid chromatography. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, v. 1072, p. 193-198, JAN 1 2018. (12/24345-4, 15/10928-6, 15/26058-0)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

DIAS, Paulo Victor Sarmento. Development of production process and characterization of periplasmic interferon-α2a expressed in Escherichia Coli. 2017. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Pesquisas Energéticas e Nucleares (IPEN/BT) São Paulo.