Production and characterization of anti-recombinant human erythropoietin (rhEPO) monoclonal antibody.
Anti-rhEPO McAb is valuable for the determination of recombinant human erythropoietin (rhEPO) levels for diagnosis of renal anemia and for doping control analysis. In this paper, anti-rhEPO hybridoma was prepared by fusion of SP2/0 murine myeloma cells with spleen cells isolated from immunized BALB/c mouse, using an enzyme-linked immunosorbent assay (ELISA) method to screen the positive hybridoma. The purified McAb was characterized by ELISA, SDS-PAGE, and Western-blotting. Experimental results showed that the subclass and the light chain of anti-rhEPO McAb was IgG1 and kappa light chain. The molecular weight of anti-rhEPO McAb was 166,000 Daltons. The affinity constant (K(aff)) of anti-rhEPO McAb with coated antigen was 5.0 x 10(5)L/mol.
Yan
Jin
J
The Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, PR China.
Wang
Shan
S
Mi
Jie-Bo
JB
Guo
Zhen-Quan
ZQ
Chang
Wen-Bao
WB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry* / Q000276:immunology; D000915:Antibody Affinity; D015153:Blotting, Western; D002459:Cell Fusion; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004921:Erythropoietin / Q000032:analysis / Q000276:immunology*; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D009101:Multiple Myeloma; D011994:Recombinant Proteins
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D004921:Erythropoietin