Development and evaluation of recombinant human erythropoietin (rhEPO) production system

Abstract

Erythropoietin (EPO) is a glycoprotein hormone primarily produced in the kidneys, playing a vital role in hematopoiesis by regulating erythroid progenitor cell proliferation and maturation in the bone marrow. Recombinant human erythropoietin (rhEPO) is widely used to treat anemia associated with chronic kidney disease, chemotherapy. However, the structure of EPO presents challenges for its production, affecting its therapeutic efficacy and half-life. To address these challenges, we developed stable rhEPO-expressing HEK293T cell lines using an inducible YFP-hEPO construct. Additionally, we engineered a Flag-hEPO to facilitate purification. The biological activity of rhEPO produced from these lines was validated by assessing its ability to activate the proliferation and EPOR signaling pathway in HUDEP-2 cells, showing comparable efficacy to commercial EPO. These findings lay a foundation for future efforts focused on enhancing rhEPO production, purification, and bioactivity, paving the way for enhanced clinical applications.