Production and characterization of monoclonal antibody against recombinant human erythropoietin.
作者信息
Mi Jie-Bo, Yan Jin, Ding Xiao-Jie, Guo Zhen-Quan, Zhao Mei-Ping, Chang Wen-Bao
机构信息
Beijing National Laboratory for Molecular Sciences (BNLMS), College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
出版信息
Biomed Environ Sci. 2007 Jun;20(3):184-8.
OBJECTIVE
To produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids.
METHODS
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.
RESULTS
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.
CONCLUSIONS
The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.
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