Erythropoietin assay in mice made polycythemic by transfusion of heterologous red cells.
A simple in vivo bioassay suitable for routine testing of quality control of recombinant human erythropoietin (rHu-EPO) analogues was developed. Mice made polycythemic by intraperitoneal injection of 1.2 ml of a 80% suspension of heterologous (rat) red cells were used as assay animals and splenic 59Fe uptake as expression of the response to rHu-EPO. The assay took three days and the following schedule is proposed: 1) intraperitoneal injection of 1.2 ml of washed packed red cells obtained from donor rats, 2) subcutaneous injection of test material 4-5 h after transfusion, 3) intravenous administration of 59Fe tracer 48 h later, and 4) determination of splenic isotope uptake 6 h after injection. This method for the in vivo bioassay of rHu-EPO analogues is an economical and reliable alternative to the existing bioassays of the hormone.
Conti
M I
MI
Cátedra de Fisiología, Facultad de Odontología, Universidad de Buenos Aires, Argentina.
Martínez
M P
MP
Barceló
A C
AC
Alippi
R M
RM
Bozzini
C E
CE
D000818:Animals; D001681:Biological Assay; D017707:Erythrocyte Transfusion; D004920:Erythropoiesis; D004921:Erythropoietin / Q000032:analysis*; D005260:Female; D007504:Iron Radioisotopes; D051379:Mice; D011086:Polycythemia / Q000209:etiology / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins
D007504:Iron Radioisotopes; D011994:Recombinant Proteins; D004921:Erythropoietin