Expression and bioactivity of recombinant canine erythropoietin.
OBJECTIVE
To produce recombinant canine erythropoietin (rcEPO) and compare its biological activity with that of recombinant human EPO (rhEPO).


METHODS
C57BL/6J mice.


METHODS
The gene encoding cEPO was isolated from a genomic library and subcloned into an eucaryotic expression vector. Production of rcEPO was achieved by stable transfection of the expression construct into Chinese hamster ovary cells. Biological activity was evaluated in vitro by analyzing the mitogenic activity of rcEPO on murine erythroid progenitor cells. In vivo bioactivity was assessed in mice by measuring the ability of rcEPO to increase blood reticulocyte counts.


RESULTS
Size and glycosylation of rcEPO expressed in Chinese hamster ovary cells were similar to values for commercial rhEPO. Canine and human EPO stimulated proliferation of murine erythroid progenitor cells in vitro and murine reticulocytosis in vivo in a dose-dependent manner.


CONCLUSIONS
Comparable biological activity was observed for rcEPO and rhEPO in the 2 murine-based assay systems studied. By avoiding interspecies variation in protein structure and the resulting potential for immunogenicity, rcEPO should represent a better option than rhEPO for treatment of dogs with erythropoietin-dependent anemia.


CONCLUSIONS
Therapeutic use of rhEPO in companion animals is limited by its immunogenicity and the resulting potential to induce pure red cell aplasia. Development and availability of species-specific EPO preparations should avoid this problem.
MacLeod
J N
JN
Department of Biomedical Sciences, James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Tetreault
J W
JW
Lorschy
K A
KA
Gu
D N
DN
D000818:Animals; D016466:CHO Cells; D002455:Cell Division; D006224:Cricetinae; D004285:Dogs; D015672:Erythroid Precursor Cells / Q000187:drug effects / Q000378:metabolism; D004921:Erythropoietin / Q000096:biosynthesis* / Q000378:metabolism; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D017701:Reticulocyte Count / Q000187:drug effects
D011994:Recombinant Proteins; D004921:Erythropoietin