CHIMERIZATION AND HUMANIZATION OF AN anti-IFN-alpha2b ANTIBODY FRAGMENT FOR THERAPEUTIC USE

ATTALLAH, C; BURTET, R; MARANHAO, A; ETCHEVERRIGARAY, M; OGGERO, M

Buenos Aires

Congreso; IV LASID Meeting, LXIII Argentinian Society for Immunology Meeting, II French Society for Immunolgy Meeting.; 2015

LASID FAIC SAI

Recently anti-IFN-alpha recombinant antibodies therapy has emerged as a strategy to treat the Systemic Lupus Erythematosus (SLE). The therapeutic antibodies are potentially immunogenic. Several approaches were employed to reduce this risk, as the generation of chimeric, humanized and fully human molecules. In this work, we generated two anti-IFN-alpha2b, chimeric and humanized scFv-Fc (single-chain variable fragment fused to Fc region), for therapeutic use. Therefore, the Fc region of human IgG1 was fused to a murine anti-IFN-alpha2b scFv, previously obtained in our lab, by polymerase chain reaction (PCR). CHO-K1 (Chinese hamster ovary), HEK293 (Human Embryonic Kidney) and NS0 (derived from murine myeloma) cells were transfected. The capacity of the fusion proteins produced in cell culture supernatant to bind the rhIFN-alpha2b was evaluated by indirect enzyme-linked immunosorbent assay (ELISA). Later the murine scFv was humanized in silico. This molecule was fused to the Fc region of the human IgG1. HEK293 and CHOK1 cells were transfected and the capacity of the protein produced in cell culture supernatant to bind the rhIFN-alpha2b was evaluated by indirect ELISA.We demonstrated the ability of both scFv-Fc molecules to recognize the rhIFN-alpha2b. These antibodies are potential therapeutic agents for SLE.