N- and O-Glycosylation Muteins of Recombinant Human Erythropoietin Secreted From BHK-21 Cells

• Published in: Blood 1995-03, Vol.85 (5), p.1229-1236
• Main Authors: Fibi, M.R., Hermentin, P., Pauly, J.U., Lauffer, L., Zettlmeissl, G.
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Binding, Competitive; Biological and medical sciences; Carbohydrate Conformation; Cell differentiation, maturation, development, hematopoiesis; Cell Line; Cell physiology; Cricetinae; Erythropoietin - analogs & derivatives; Erythropoietin - metabolism; Fundamental and applied biological sciences. Psychology; Glycosylation; Humans; Kidney; Mesocricetus; Molecular and cellular biology; Molecular Sequence Data; Mutagenesis, Site-Directed; Protein Processing, Post-Translational; Receptors, Erythropoietin - metabolism; Recombinant Fusion Proteins - metabolism; Secretory Rate; Structure-Activity Relationship
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V85.5.1229.bloodjournal8551229

Single-site glycomuteins of recombinant human erythropoietin (rhuEpo) were constructed and transiently and stably expressed in BHK-21 cells. The transient expression levels varied among muteins, being highest for mutein rhuEpoGln24 followed by wild-type rhuEpo (rhuEpowt). All other glycomuteins, including rhuEpoGln38, rhuEpoGln83, rhuEpoThr126, and rhuEpoGly126, were secreted at lower levels than rhuEpowt. Muteins expressed in stable cell lines showed similar differences in expression levels. Also each mutein could be affinity-purified from culture supernatants, and was biologically active in vivo. Based on secretion rates from BHK-21 cells, the most potent erythropoietin was rhuEpoGln24. This mutein is also considered to have biologic activities that are superior to rhuEpowt.