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

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)