Abbas Farhadian, Hooman Kaghazian, Mohammad Reza Akbari Eidghahi

Abstract

For producing recombinant erythropoietin, several cell lines are used. More common, CHO cell lines are employed to manufacture this protein. So, large amounts of impurities are produced during the process. Therefore, chromatography methods are used for purification. Optimizing the chromatography condition can improve the production process. Initially, the harvest was collected from cultured CHO cell lines. Then, the sample was applied to gel filtration, IEX, and high-resolution chromatography with different buffering conditions. The amount of obtained protein was measured by ELISA. SDS-PAGE, Capillary Zone Electrophoresis method, and peptide map were used to determine the effect of buffer condition on the eluted protein. The results indicated that the buffering condition of chromatography columns affected the quality and quantity of obtained proteins. Molecular weight and peptide maps of all samples were similar to the standard sample. CZE test showed that all samples were the same as the standard sample, except for one case. The results indicated the least change in chromatography conditions highly impresses our obtained proteins and led to significant variation in their quality and quantity‎.