IBYME   02675
INSTITUTO DE BIOLOGIA Y MEDICINA EXPERIMENTAL
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Production of anti-IFN-a2b monoclonal antibody in microfluidic devices.
Autor/es:
BOURGUIGNON, NATALIA; PEÑAHERRERA, ANA; LERNER, BETIANA; KARP, PAOLA; LASORSA, CARLOS; PEREZ, MAXIMILIANO; ATTALLAH, CAROLINA; PAYÉS, CRISTIAN; HELGUERA, GUSTAVO
Lugar:
Buenos Aires
Reunión:
Congreso; III International Congress in Translational Medicine; 2016
Institución organizadora:
International Master Program in Biomedical Sciences (IMBS)
Resumen:
Monoclonal antibodies production for therapeutic use is an expanding area of the biopharmaceutical industry. The traditional production in bioreactors faces challenges related with the quality of product, glycosylation control and reproducibility. One approach to improve the control is to reduce the scale of the production platform through microdevices (1). In our laboratory we are developing a microfluidic device to produce therapeutic biomolecules. Recently anti-IFN-alpha recombinant antibodies has emerged as a strategy to treat the Systemic Lupus Erythematosus (SLE). In this work, we evaluated the production in this device of an anti-IFN-a2b recombinant minibody consisting in the variable region of a murine monoclonal antibody fused genetically via a flexible linkers to a Fc region from a human origin. This recombinant antibody show neutralizing ability of antiviral and antiproliferative activity of IFN-a2b, in vitro and has potential therapeutic use to treat Systemic Lupus Erythematosus (SLE) (2). In this study, two cell lines that produce the antibody, CHO-K1 (Chinese hamster ovary) and HEK293 (Human Embryonic Kidney) were cultured in poly-dimetil silaxane (PDMS) microfluidic devices with architecture of four microchannels with cisterns. The microdevices with wide channels were covered with poly-lysine to favor the attachment of cells to the bottom, which benefits the cell expansion in these devices (3). Cells were able to grow, reaching confluency and covering the total area of the microchip after 8 days of incubation. At this stage we began the antibodies production analysis and the productivity of anti-IFN-a2b in CHO-K1 and HEK293 was monitored for 18 days by binding determination to rhIFN-alpha2b using indirect ELISA. An increase in antibody production was observed over time reaching values of 58,3 ug/ml for CHO-K1 and 24,0 ug/ml for HEK293 cells at the last day of incubation. Production achieved in the microchip was higher than that obtained in T-flasks, probably due its high ratio of area covered by cells / device surface. We demonstrated for the first the successful growth and production of anti-IFN-a2b monoclonal antibodies by cells cultured in microfluidic devices.