Chinese Journal of Chromatography

Chinese Journal of Chromatography

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Purification of Plasmid DNA Using Anion-Exchange Chromatography and Removal of Endotoxin

PI Wenhui1,2, SUN Congjian3, SONG Zhiqiang2, MA Lianying3, LIU Shuping3, BAI Dingping2, CAI Yidong3, LIU Shouren2   

  1. 1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China; 2.Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China; 3.Xinjiang Tianwu Science & Technology Development Co., Ltd., Urumqi 830011, China
  • Received:2007-05-21 Revised:2007-07-18 Online:2007-11-30 Published:1984-12-25

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Abstract:

The plasmid DNA was purified using the Fractogel EMD TMAE (M) anion exchange media. The dynamic loading capacity of this media for plasmid DNA was 0.62 mg/mL. It was found that when the lysate of bacteria was incubated with either Triton X-114 or Triton X-100 before purification, the average content of endotoxin left in the purified plasmid DNA was 6.42 EU/mg or 9.50 EU/mg, respectively. These results were much lower than that without pre-incubation (67.82 EU/mg). The purification of plasmid DNA and the removal of endotoxin were achieved in one step using anion-exchange chromatography. This method is simple, fast and of low cost.

Key words: Triton X-114, endotoxin, plasmid DNA, purification, removal , Triton X-100, anion-exchange chromatography