Boratyński Janusz, Szermer-Olearnik Bożena
Laboratory of Biomedical Chemistry - "Neolek", Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, ul. Rudolfa Weigla 12, Wroclaw, PL-53-114, Poland.
Methods Mol Biol. 2017;1600:107-112. doi: 10.1007/978-1-4939-6958-6_10.
Lipopolysaccharide (LPS, endotoxin, pyrogen) which is a component of the outer membrane of most Gram-negative bacteria is a troubling contaminant of crude bacteriophage suspension. Therefore, its removal is important for bacteriophage applications especially in preparations dedicated for use in therapy with bacterial infections treatment. The method presented here is used for extractive removal of endotoxins from bacteriophage preparations with a water immiscible solvent such as 1-octanol. During extraction most of the phage lytic activity is retained in the aqueous phase, while endotoxin accumulates in the organic solvent. The levels of endotoxin in the aqueous bacteriophage rich fraction are determine by Limulus Amebocyte Lysate or EndoLISA assay and are extremely low.
脂多糖（LPS，内毒素，热原）是大多数革兰氏阴性菌外膜的组成成分，是粗制噬菌体悬浮液中令人困扰的污染物。因此，去除脂多糖对于噬菌体的应用非常重要，特别是在专门用于治疗细菌感染的制剂中。本文介绍的方法用于用与水不混溶的溶剂（如1-辛醇）从噬菌体制剂中萃取去除内毒素。在萃取过程中，大部分噬菌体裂解活性保留在水相中，而内毒素则积聚在有机溶剂中。富含噬菌体的水相部分中的内毒素水平通过鲎试剂或内毒素酶联免疫吸附测定法测定，结果极低。
