Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Purification of Filamentous Bacteriophage M13 by Expanded Bed Anion Exchange Chromatography

  • Tau Chuan (Department of Process and Food Engineering, Faculty of Engineering) ;
  • Chee Kin (Department of Process and Food Engineering, Faculty of Engineering) ;
  • Wen Siang (Department of Biochemistry and Microbiology, Faculty of Science and Environmental Studies) ;
  • Beng Ti (Department of Chemical and Environmental Engineering, Faculty of Engineering) ;
  • Wan, Wan-Mohammad (Department of Process and Food Engineering, Faculty of Engineering) ;
  • Arbakariya (Department of Biotechnology, Faculty of Food Science and Biotechnology, Universiti Putra Malaysia)
  • Published : 2004.09.01
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Abstract

In this paper, we investigated the development of a simplified and rapid primary capture step for the recovery of M13 bacteriophage from particulate-containing feedstock. M13 bacteriophage, carrying an insert, was propagated and subsequently purified by the application of both conventional multiple steps and expanded bed anion exchange chromatography. In the conventional method, precipitation was conducted with PEG/NaCl, and centrifugation was also performed. In the single step expanded bed anion exchange adsorption, UpFront FastLine$\_$TM/20 (20mm i.d.) from UpFront Chromatography was used as the contactor, while 54$m\ell$ (H$\_$o/=15cm) of STREAMLINE DEAE (p=1.2 g/㎤) from Amersham Pharmacia Biotechnology was used as the anion exchanger. The performance of the two methods were evaluated, analysed, and compared. It was demonstrated that the purification of the M13 bacteriophage, using expanded bed anion exchange adsorption, yielded the higher recovery percentage, at 82.86％. The conventional multiple step method yielded the lower recovery percentage, 36.07％. The generic application of this integrated technique has also been assessed.

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References

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