Production and characterization of monoclonal antibody against recombinant human erythropoietin / 生物医学与环境科学（英文）

ABSTRACT

<p><b>OBJECTIVE</b>To produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids.</p><p><b>METHODS</b>rHuEPO was covalently coupled with bovine serum albumin (BSA) and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western blot.</p><p><b>RESULTS</b>The isotype of F3-mAb was found to be IgM with an affinity constant of 2.1 x 10(8) L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work.</p><p><b>CONCLUSIONS</b>The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.</p>