Biphasic Cultivation for Inhibition of Sialidase from CHO Cells Producing Albumin-erythropoietin

초록

Chinese hamster ovary (CHO) cells are capable of producing recombinant proteins with human-like glycosylation profile. Nglycosylation of pharmaceutical proteins influences on their half-life, immunogenicity, and efficacy. Especially, terminal sialic acid residues of N-linked glycan have a profound impact on the circulatory half-life of glycoproteins as they prevent the recognition by asialo-glycoprotein receptor. For that reason, it is important to maximize sialic acid contents of glycoprotein to increase half-life. Desialylation of recombinant glycoprotein in cell culture media is predominantly caused by the release of intracellular sialidase by cell lysis. It also lowers in vivo sustainability of recombinant glycoproteins. Sialidase activity is known to be optimal at 37℃ and pH 5.9. In this study, biphasic culture of recombinant CHO cells producing a fusion protein, albumin-erythropoietin (Alb-EPO), was investigated. Sialidase activity could be reduced by shifting culture conditions such as pH and temperature. In conclusion, this biphasic cultivation method was found to improve the quality of therapeutic proteins.