以TritonX-114液相分离法去除质粒溶液中的内毒素

Removal of Endotoxin from Plasmid Solutions by Triton X-114 Phase Separation

目的:采用Triton X-114液相分离法去除质粒溶液中的内毒素,以保证实验动物的安全和结果的准确性。方法:通过碱裂解法提取质粒pVAX1和pVAX1-hLDHC,用聚乙二醇6000沉淀对质粒进一步纯化;用Triton X-114抽提的方法,去除质粒溶液中的内毒素。结果:通过3轮Triton X-114抽提,能够将质粒溶液中内毒素水平降至1.95 EU/mL,质粒样品的回收率为79.8%,质量保持不变。结论:Triton X-114液相分离法是一种非常有效的去除质粒溶液中内毒素的方法。

Objective： To remove the endotoxin from plasmid solutions by Triton X-114 phase separation for the safety of the experimental animals and the accurate of the resultes. Methods： The plasmids pVAX1 and pVAXI-hLDHC were isolated by alkaline lysis and polyethylene glycol precipitation. Endotoxin in plasmid solution was removed by three rounds of Triton X-114 phase separation. Results： The endotoxin level in plasmid solution was reduced to 1.95 EU/mL after the third extraction, and the recovery ratio of plasmid was about 79.8%. No changed in quanlity of the plasmid was observed. Conclusion： The Triton X-114 phase separation was a very effective method for removing endotoxin from plasmid solutions.