Production of Recombinant Woodchuck IFN alfa and Development of Monoclonal Antibodies
Pedro Berraondo, Julien Crettaz, Laura Ochoa, Africa Vales, Juan Ruiz, Jesús Prieto, Eduardo Martinez-Ansó, and Gloria González-Aseguinolaza
Laboratory of Gene Therapy of Viral Hepatitis, Division of Hepatology and Gene Therapy, Center for Applied Medical Research (CIMA), Clínica Universitaria/School of Medicine, University of Navarra, Pamplona, Spain
Interferon alphas the first line treatment for chronic hepatitis B and C. In order to test new; delivery systems and investigate the function of this cytokine in the woodchuck model, the best animal model of chronic hepatitis B, we produced and purified recombinant woodchuck; and used it to produce monoclonal antibodies.
IFN was cloned in a prokaryotic expression system, expressed as His-tagged protein and then purified. The IFN protein was found to induce STAT-3 phosphorylation, to enhance IFN -oligoadenylate synthetase mRNA levels and to possess a potent antiviral activity. Two monoclonal antibodies were obtained through immunization of rats with rwIFN?5. Both recognized IFN in western blot analysis and one was able to neutralize the antiviral activity of the IFN and lymphoblastoid IFN? preparations. Finally, a capture IFN ELISA was developed using both antibodies.
In summary, the tools generated in this study will allow different forms of IFN delivery as well as different combination therapies in woodchuck hepatitis virus infection to be tested, thus providing useful information for the design of new strategies to treat chronic hepatitis B in humans.
CITA DEL ARTÍCULO J Interferon Cytokine Res. 2009 Feb;29(2):75-82