质粒DNA的阴离子交换色谱法纯化及内毒素去除

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

采用Fractogel(EMD TMAE(M)强阴离子交换介质分离纯化质粒脱氧核糖核酸(DNA),该介质对质粒DNA的动态载样量达0.62mg/mL。用Triton X-114或Triton X-100预先处理质粒DNA的裂解澄清液后,经阴离子交换色谱分离纯化获得的质粒DNA中内毒素含量分别为6.42EU/mg和9.50EU/mg,显著低于未经Triton处理的裂解澄清液(67.82EU/mg)。该法实现了阴离子交换色谱一步纯化质粒DNA的目的,具有简便、省时、成本低等特点。

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 0.52 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 5.42 EU/mg or 9.50 EU/mg, respectively. These results were much lower than that without pre-incubation （57.82 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.