OPTIMAL SDS-PAGE AND WESTERN BLOT TECHNIQUES FOR IDENTIFICATION TESTING OF ERYTHROPOIETIN IN HIGH-TECH BIOLOGICAL PRODUCT

Abstract

Erythropoietin (EPO) is an erythropoiesis-stimulating factor produced by recombinant DNA technology, used in the treatment of anemia. Identification testing using sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western Blot are important quality control procedures for biological products containing EPO. In National institute for control of vaccine and biologicals (NICVB), our researcher established an EPO identification approach using SDS-PAGE and Western Blot techniques on two biological products containing epoietin alpha. At the same time, the optimal primary and secondary antibody concentrations were selected for nitrocellulose membrane hybridization testing using two dilutions of 1:500 and 1:800. In addition, 3,3',5,5'-tetramethylbenzidine (TMB) was used to detect the antigen-antibody reaction. The results showed that using TMB as the detection substrate provided quick results, simple preparation and an optimal implementation process.  Our results provide evidence to implement SDS-PAGE and Western Blot for identification of EPO in High Tech Biological Products in NICVB