Post-translational modification of a chimeric EPO-Fc hormone is more important than its molecular size in defining its in vivo hematopoietic activity

摘要: Abstract Background Recombinant erythropoietin (EPO) has been marketed as biopharmaceutical for anemia and chronic renal failure. Long-acting EPO variants that aimed at achieving less frequent dosing have generated, either by the addition of glycosylation sites or increasing its molecular weight. Methods The hEPO cDNA linked to human IgG Fc fragment was cloned a single codifying gene on pAdtrack-CMV vector, yielding recombinant adenoviral genome. For in vitro vivo expression assays cervical cancer cell line (SiHa) nulliparous goats were used, respectively. hematopoietic activity EPO-Fc, expressed differential increment hematocrit evaluated B6D2F1 mice. NP-HPLC 2AB-labeled N-glycan carried out profile analysis. Results direct transduction mammary secretory cells with vector is robust methodology obtain high levels up 3.5 mg/mL goat's milk. SiHa-derived EPO-Fc showed significant improvement compared commercial counterpart homologous milk-derived EPO-Fc. role weight seemed be important enhancing However, lack sialylated multi-antennary resulted lower biological activity. Conclusions low content tri- tetra-antennary N-glycans chimeric hormone, goat gland epithelial cells, defined General significance plays more than increased