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Article in English | WPRIM (Western Pacific) | ID: wpr-249868

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>


Subject(s)
Animals , Female , Humans , Mice , Antibodies, Monoclonal , Allergy and Immunology , Antibody Affinity , Dose-Response Relationship, Drug , Enzyme-Linked Immunosorbent Assay , Erythropoietin , Allergy and Immunology , Immunoglobulin G , Allergy and Immunology , Immunoglobulin M , Allergy and Immunology , Mice, Inbred BALB C , Molecular Weight , Recombinant Proteins
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