DNA removal on different chromatography supports used for EPO purification by spike studies |
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| Authors: | L. Hernández A. Rodríguez V. Peña L. Zumalacárregui D. Amaro |
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| Affiliation: | 1. Development Product Department, Manufacturing Plant of Recombinant Proteins and Vaccines, Center for Molecular Immunology, Playa, Havana, Cuba;2. Department of Chemical Engineering, Faculty of Chemical Engineering, Higher Polytechnic Institute José Antonio Echeverría, Marianao, Havana, Cubalourdes@cim.sld.cu;4. Department of Chemical Engineering, Faculty of Chemical Engineering, Higher Polytechnic Institute José Antonio Echeverría, Marianao, Havana, Cuba;5. Department of Chemical Engineering, Faculty of Chemical Engineering, Higher Polytechnic Institute José Antonio Echeverría, Marianao, Havana, Cuba |
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| Abstract: | Chromatographic techniques are used in the purification step of human recombinant erythropoietin production process to obtain a reliable product with high purity. Anion-exchange chromatography supports have proved high efficient in removing contaminants such as DNA. For that reason, the DNA removal was determined by spike studies, on three anion-exchange chromatographic supports: gel, membrane, and monolithic column, which is used in intermediate purification stage. This study showed that membrane and monolith columns have very good results in the removal of contaminants at this step. Log removal values (LRV) greater than 3.5 were obtained from DNA spike clearance studies. Monolithic column was determined as the best technological proposal, with more than 4 LRV, 7.72?mg DNA per milliliter of adsorbent and 85% protein recovery in nonspike run. The results of this study may be used as a guide in the selection of commercially available chromatography supports for intermediate purification steps in recombinant protein production. |
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| Keywords: | Chromatography supports DNA removal EPO purification |
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