N- and O-glycosylation muteins of recombinant human erythropoietin secreted from BHK-21 cells.
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.
Research Laboratories of Behringwerke AG, Department of Preclinical Research of Therapeutics, Marburg, Germany.
M R
MR
Fibi
P
P
Hermentin
J U
JU
Pauly
L
L
Lauffer
G
G
Zettlmeissl
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D002236:Carbohydrate Conformation; D002460:Cell Line; D006224:Cricetinae; D004921:Erythropoietin / Q000031:analogs & derivatives* / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D007668:Kidney; D008647:Mesocricetus; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011499:Protein Processing, Post-Translational; D017467:Receptors, Erythropoietin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012636:Secretory Rate; D013329:Structure-Activity Relationship
D017467:Receptors, Erythropoietin; D011993:Recombinant Fusion Proteins; D004921:Erythropoietin