Purification and concentration of mycobacteriophage D29 using monolithic chromatographic columns.

Keyang Liu, Zhanbo Wen, Na Li, Wenhui Yang, Lingfei Hu, Jie Wang, Zhe Yin, Xiaokai Dong, Jinsong Li

State Research Center for Bio protective Equipment & Engineering Technology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China.

Journal of virological methods 2012 Dec

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Bacteriophages are used widely in many fields, and phages with high purity and infectivity are required. Convective interaction media (CIM) methacrylate monoliths were used for the purification of mycobacteriophage D29. The lytic phages D29 from bacterial lysate were purified primarily by polyethylene glycol 8000 or ammonium sulphate, and then the resulting phages were passed through the CIM monolithic columns for further purification. After the whole purification process, more than 99% of the total proteins were removed irrespective which primary purification method was used. The total recovery rates of viable phages were around 10-30%. Comparable results were obtained when the purification method was scaled-up from a 0.34 mL CIM DEAE (diethylamine) monolithic disk to an 8 mL CIM DEAE monolithic column. Copyright © 2012 Elsevier B.V. All rights reserved.

Citation

Keyang Liu, Zhanbo Wen, Na Li, Wenhui Yang, Lingfei Hu, Jie Wang, Zhe Yin, Xiaokai Dong, Jinsong Li. Purification and concentration of mycobacteriophage D29 using monolithic chromatographic columns. Journal of virological methods. 2012 Dec;186(1-2):7-13