Characterization of an anti-interferon-alpha2b chimeric fusion protein for therapeutic use

ATTALLAH, C; MUFARREGE, E; ETCHEVERRIGARAY, M; OGGERO, M

Mar del Plata

Congreso; LXIV Reunión anual de la sociedad argentina de inmunología; 2016

SAIC - SAI - SAFE

Since the late 70?s, the interferon alpha (IFN-alpha) has been associated with systemic lupus erythematosus (SLE). From that moment, an impressive number of studies have confirmed the importance of IFN-alpha in lupus, and have prompted investigation of anti-IFN-alpha therapeutic strategies. In this work, we have characterized an anti-IFN-alpha2b chimeric fusion protein in terms of its IFN-alpha affinity, in vitro biological activity and the ability to produce antibodies in transgenic animals inoculated with this protein produced from different types of cells. Therefore, CHO-K1 (Chinese hamster ovary), HEK293 (Human Embryonic Kidney) and NS0 (derived from murine myeloma) cells were used to produce the fusion protein. The IFN-alpha affinity is in the order of 10-8 M-1. The fusion proteins produced for different types of cells presented the ability to neutralize the IFN-alpha antiproliferative and antiviral activities. On the other hand, the proteins were stable until 10 min at 55°C, preserving the ability of binding the IFN-alpha2b. In addition, we evaluated the immunogenicity at humoral level by in vivo assays in HLA-DR1 transgenic mice. These animals were inoculated with fusion proteins and we evaluated the anti-anti-IFN-alpha2b antibodies production. The highest antibodies title was obtained with the protein derived from HEK293 cells. This characterization allows us take into account the abilities of an anti-IFN-alpha2b antibody as potential therapeutic agent for SLE and the cell types to produce it.