Cotten M, Baker A, Saltik M, Wagner E, Buschle M
IMP, Research Institute of Molecular Pathology, Vienna, Austria.
Gene Ther. 1994 Jul;1(4):239-46.
Endotoxin (lipopolysaccharide, LPS) is commonly found as a contaminant in plasmid DNA preparations. We demonstrate here that the quantities of LPS typically contaminating DNA preparations can generate a toxicity to primary cells (primary human skin fibroblasts, primary human melanoma cells) in the presence of entry-competent adenovirus particles. Toxicity can be observed with as little as 100 ng/ml free LPS or 100 pg/ml LPS when the LPS is assembled into polylysine/adenovirus complexes. Simple and effective methods of removing the contaminating LPS using either a polymyxin B resin or Triton X-114 extraction are described. Treatment of DNA samples to remove LPS eliminates the toxicity to primary cells.
内毒素（脂多糖，LPS）在质粒DNA制备物中通常作为污染物被发现。我们在此证明，在具有感染能力的腺病毒颗粒存在的情况下，通常污染DNA制备物的LPS量可对原代细胞（原代人皮肤成纤维细胞、原代人黑色素瘤细胞）产生毒性。当LPS组装成聚赖氨酸/腺病毒复合物时，低至100 ng/ml游离LPS或100 pg/ml LPS即可观察到毒性。描述了使用多粘菌素B树脂或Triton X-114萃取去除污染LPS的简单有效方法。处理DNA样品以去除LPS可消除对原代细胞的毒性。
