Plasticity of the HEK-293 cells, related to the culture media, as platform to produce a subunit vaccine against classical swine fever virus

Abstract

Abstract Classical swine fever (CSF) is a contagious disease that causes a high mortality to domestic and wild pigs. Its causative agent is an enveloped Pestivirus named Classical Swine Fever Virus (CSFV). Due to the huge economic affectations produced by this disease to porcine industry, several vaccines have been developed using principally the CSFV E2 glycoprotein. Recently, a subunit vaccine based on this structural protein of the CSFV fused to the porcine CD154 molecule as immunomodulator named E2-CD154 was assayed by us. This chimeric protein was produced in the Human Embryonic Kidney (HEK-293) cell line. In this work, the growth and the expression profiles of HEK-293 E2-CD154 cells in four commercially available culture media were studied. The oligosaccharide structures in the N-glycosylation patterns of the E2-CD154 protein produced by this cell line in 10 L fermenters with two different culture media were also analyzed. In addition, the neutralizing antibody response generated in mice vaccinated with these antigens was assayed. Our results suggest that the culture media CDM4HEK293 and SFM4HEK293 which are recommended for HEK-293 growth are the best choice to growth the cell clone expressing the E2-CD154 protein. The glycosylation pattern and the neutralizing antibody response generated by the E2-CD154 protein were independent of the culture medium used which demonstrates the high reproducibility and consistency among protein batches produced by HEK-293 cells even in different culture conditions.

Keywords

• Glycoprotein E2-CD154
• HEK-293 cells
• Culture media
• Growth profiles
• N-Glycosylation