Purification of filamentous bacteriophage M13 by expanded bed anion exchange chromatography

Chuan Ling Tau, Kin Loong Chee, Siang Tan Wen, Ti Tey Beng, Wan Mohammad Wan Abdullah, Arbakariya Ariff

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15 Citations (Scopus)

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™20 (20 mm i.d.) from UpFront Chromatography was used as the contactor, while 54 ml (Ho=15cm) of STREAMLINE DEAE (ρ=1.2 g/cm3) 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.

Original languageEnglish
Pages (from-to)228-232
Number of pages5
JournalThe Journal of Microbiology
Volume42
Issue number3
Publication statusPublished - Sep 2004
Externally publishedYes

Keywords

  • Adsorption
  • Anion exchanger
  • Bacteriophage M13
  • Expanded bed chromatography
  • Protein purification

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