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Dimeric erythropoietin fusion protein with enhanced erythropoietic activity in vitro and in vivo by Bruno Dalle, Annie Henri, Philippe Rouyer-Fessard, Mickaël Bettan, Daniel Scherman, Yves Beuzard, and Emmanuel Payen Blood Volume 97(12):3776-3782 June 15, 2001 ©2001 by American Society of Hematology The linker fragment used to bridge Epo encodes a peptide of 9 residues.When annealed, the 2 oligonucleotides shown in this figure present protruding BamHI and EcoRI extremities. Bruno Dalle et al. Blood 2001;97:3776-3782 ©2001 by American Society of Hematology ELISA overestimates dimeric erythropoietin by 5 times.(Lane 1) 3 U, (lane 2) 2 U, and (lane 3) 1 U human HA-tagged dimeric erythropoietin and 0.6 U (lane 4), 0.4 U (lane 5), and 0.2 U (lane 6) human HA-tagged monomeric erythropoietin (as measured by R&D ELI... Bruno Dalle et al. Blood 2001;97:3776-3782 ©2001 by American Society of Hematology Dimeric erythropoietin induces higher hematocrit increase than monomer.Two groups of 6 C57BL/6 mice were injected intraperitoneally with 3 × 300 U/kg (as determined by proliferation assay) of hEpoD (A) and hEpoM (B) on days 1, 3, and 5. Bruno Dalle et al. Blood 2001;97:3776-3782 ©2001 by American Society of Hematology Dimeric erythropoietin is 6 times more efficient in vivo than monomer.Six groups of 6 C57BL/6 mice were injected intraperitoneally, either with 3 times sodium chloride or with 3 times 300 U/kg, 900 U/kg, or 1800 U/kg monomeric hEpo (M) or with 3 times 300 U... Bruno Dalle et al. Blood 2001;97:3776-3782 ©2001 by American Society of Hematology Human dimeric and monomeric forms of erythropoietin have the same pharmacokinetics in vivo.Six groups of 3 C57BL/6 mice were injected intravenously (A), intraperitoneally (B), or subcutaneously (C) with 200 U/kg (as determined by proliferation assay) hEpoM ... Bruno Dalle et al. Blood 2001;97:3776-3782 ©2001 by American Society of Hematology Human dimeric erythropoietin has enhanced erythropoietic activity.CD34+ cells were purified and grown in semisolid medium containing 0.1 to 0.6 U/mL (as determined by proliferation assay) hEpoM (black) or hEpoD (white). Bruno Dalle et al. Blood 2001;97:3776-3782 ©2001 by American Society of Hematology Human dimeric erythropoietin stimulates late erythroid progenitors.CD36+ cells were purified and grown in semisolid medium containing 0.1 to 0.4 U/mL (as determined by proliferation assay) hEpoM (black) or hEpoD (white). Bruno Dalle et al. Blood 2001;97:3776-3782 ©2001 by American Society of Hematology Controlled secretion of dimeric erythropoietin does not induce antibodies.Four β-thalassemic mice were injected with a mix of 2 μg ptet-Off and 20 μg ptet-mEpoD. Bruno Dalle et al. Blood 2001;97:3776-3782 ©2001 by American Society of Hematology
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