GSK Vaccines Institute for Global Health, Siena, Italy.
Methods Mol Biol. 2021;2183:267-304. doi: 10.1007/978-1-0716-0795-4_14.
A variety of bacterial infections have been tackled by glycoconjugates over the recent years, and more vaccines are either under development at preclinical level or in clinical trials. So far, licensed glycoconjugate vaccines have made use of capsular polysaccharides or derived fragments. Today, many glycoconjugates are making use of other classes of sugars, in particular, the O-antigen portion of lipopolysaccharide molecules. Here, we report a simplified method for O-antigen extraction and purification that avoids the step of lipopolysaccharide isolation. Also, a selective chemistry for terminal linkage of O-antigen chains to a carrier protein is described, together with analytical methods for intermediates and final conjugate characterization.
近年来,糖缀合物在应对各种细菌性感染方面发挥了重要作用,更多的疫苗正在临床前或临床试验阶段开发中。迄今为止,已获得许可的糖缀合物疫苗利用了荚膜多糖或其衍生片段。如今,许多糖缀合物开始利用其他类型的糖,特别是脂多糖分子的 O-抗原部分。在这里,我们报告了一种简化的 O-抗原提取和纯化方法,避免了脂多糖分离的步骤。此外,还描述了一种 O-抗原链与载体蛋白末端连接的选择性化学方法,以及中间产物和最终缀合物的分析方法。