Preparation, Characterization and Application of Monoclonal Antibodies Against Human Erythropoietin
人红细胞生成素单克隆抗体的制备、鉴定及应用研究

Two monoclonal antibodies (Mabs) against hEPO are developed. We used rhEPO as antigen for immunization. The spleen cells of the immunized mice were isolated and fused with myeloma cells X63Ag8. 653. Hybridoma cell supernatant were screened for the production of anti-hEPO Mabs with ELISA, using purified rhEPO absorbed to 96-well PVC plates. One Mab is IgGl with Kd of 5. 53x10-10mol/L, while the other is IgG2b with Kd of 1. 34 x 10-10mol/L. Both Mabs can recognize rhEPO and human urine EPO of patients with aplastic anemia. They are proved to be specific to rhEPO by ELISA and Western- blot analysis. They can be used as ligands of immunoaffinity chromatography for the purification of hEPO from urine of aplastic anemia patients or the supernatant media of the transfected mammalian cells.