An ELISA specific for murine erythropoietin

Abstract

Murine recombinant erythropoietin (EPO) was purified from an EPO- producing cell line and used for the production of polyclonal monospecific anti-murine EPO antibodies in rabbits. The anti-mouse EPO antibodies were purified by two affinity chromatography procedures. In order to obtain the most sensitive ELISA, different antibody combinations were tested in the ELISA sandwich assay. The best combination was achieved with an anti-human EPO antibody as coating and the biotinylated anti-murine EPO antibody as detecting antibody. With this sandwich-ELISA a sensitive standard curve in the range of 0.6-30 mU/ml could be established. The assay provides a sensitive and reliable measure of murine EPO in serum and cell culture supernatants ranging from normal to highly elevated EPO levels.