Investigation on the removal of Depth Filter for an effective Human Recombinant Erythropoietin Production

Abstract

Erythropoietin is a glycoprotein hormone produced by the kidney, which triggers the formation of red blood cells in the bone marrow. However, due to chronic kidney disease, the erythropoietin hormone is insufficiently produced in anemic conditions. Hence, a therapeutic recombinant human erythropoietin (rhEPO) is the alternative way to overcome chronic anemia. Genetically engineered host cells, in this case, CHO cells, are used to culture and express the rhEPO. However, impurities such as host cell protein (HCP) and host cell DNA (HCD) occur during the cell culture and must be separated. The objective of this study was to investigate the role of depth filters regarding HCP and HCD elimination, as well as protein yield and purity. Depth filter is a purification technique composed of filtration and adsorption processes. Nevertheless, it may bind rhEPO to the membrane leading to a decrease in rhEPO content. During the downstream process, experiments were performed by purifying the culture with and without a depth filter. Based on the result, the depth filter plays a crucial role in removing HCP with a pI of 3.7 to 12. Depth filter is able to bind molecules with negative and positively charged due to its components. However, for HCD removal, the experiment showed an insignificant difference between using and not using depth filters. These results provide new insight into other up-scale downstream processing and performance characteristics of depth filters containing diatomaceous earth.