Production and characterization of monoclonal antibody against recombinant human erythropoietin.
- Author:
Jie-Bo MI
1
;
Jin YAN
;
Xiao-Jie DING
;
Zhen-Quan GUO
;
Mei-Ping ZHAO
;
Wen-Bao CHANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; biosynthesis; immunology; isolation & purification; Antibody Affinity; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Erythropoietin; immunology; Female; Humans; Immunoglobulin G; immunology; Immunoglobulin M; immunology; Mice; Mice, Inbred BALB C; Molecular Weight; Recombinant Proteins
- From: Biomedical and Environmental Sciences 2007;20(3):184-188
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids.
METHODSrHuEPO 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.
RESULTSThe 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.
CONCLUSIONSThe modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.
