Biology and Genetic of the Bacterial Cell Wall Unit, Innate Immunity and Leptospira Group, Institut Pasteur, Paris, France.
Methods Mol Biol. 2020;2134:53-65. doi: 10.1007/978-1-0716-0459-5_6.
Leptospira species are one of the few spirochetes to possess a lipopolysaccharide (LPS) embedded in their outer membrane. Two protocols are currently available to extract and/or purify the leptospiral lipopolysaccharides: the rapid proteinase K method and the classical hot water/phenol extraction. The first method allows to get a quick overview of the LPS O antigen structure, whereas the second method is fitted to study the immunological properties of the leptospiral LPS. These two methods will be detailed in this chapter. Methodologies to assess the quality of the purification, such as the modified silver staining coloration, will also be reviewed. Both advantages and limitations of the different analyses will be described.
钩端螺旋体属中的某些种具有包埋在外膜中的脂多糖(LPS)。目前有两种提取和/或纯化钩端螺旋体脂多糖的方案:快速蛋白酶 K 法和经典的热水/苯酚提取法。第一种方法可以快速概述 LPS O 抗原结构,而第二种方法则适合研究钩端螺旋体 LPS 的免疫学特性。本章将详细介绍这两种方法。还将回顾评估纯化质量的方法,如改良的银染色着色。将描述不同分析的优缺点。