A Reporter Gene Assay For The Bioactivity Determination Of Recombinant Human Erythropoietin

Recombinant human erythropoietin can stimulate the proliferation, differentiation andmaturation of red blood cells, is mainly used for the treatment of renal anemia. Currentauthoritative method for detecting its biological activity is the reticulocyte countingmethod which is complicated and time-consuming. Reporter gene assay has manyadvantages, simple operation, short cycle, good specificity, low degree of variation andhas become a hotspot in the research field of biological activity detection of drugs.The purpose of this study was to establish a reporter gene assay to detectrecombinant human erythropoietin biological activity rapidly. A reporter gene assay wasdeveloped based on a UT-7cell line stably transfected with luciferase gene under thecontrol of SIE，GRR，GAS. Subsequently preliminary validation of the new assay wasconducted including precision, accuracy and specificity. Then cells at three differentpassages were tested to verify the stability of the cell line. Finally, agreement betweenthe new assay and erythrometry was evaluated by Bland-Altman analysis.The new reporter gene assay showed good specificity, precision, accuracy androbustness. It was simple to operate, animal-excluded, and highly consistency witherythrometry in results. Our study demonstrated the new reporter gene assay could be aviable alternative to the traditional erythrometry.