A high-quality and cost-effective purification procedure is one of the most important requirements for manufacturing glycoconjugate vaccines. The goal of the present work was to devise a method for removing impurities such as protein and nucleic acid from Streptococcus pneumoniae serotype 2 capsular polysaccharides (CPS). The use of hydrogen peroxide for the reduction of impurities of crude CPS was investigated. Centrifugation followed by filtration decreased protein contaminant of the hydrogen peroxide-treated CPS to meet the limit specified by WHO. The nucleic acid impurity remaining was removed by a further step of endonuclease treatment to yield the purified CPS. Characterization of purified CPS was evaluated by various analytical techniques including ¹H NMR and antigenicity by competitive inhibition assay. Various hydrogen peroxide concentrations have significant impact on the antigenic property of CPS. Whereas, optimum process conditions can preserve the native characteristics of CPS.

高质量和具有成本效益的纯化程序是制造糖缀合物疫苗的最重要要求之一。本工作的目标是设计一种从肺炎链球菌血清型 2 荚膜多糖 (CPS)中去除蛋白质和核酸等杂质的方法。研究了使用过氧化氢减少粗 CPS 中的杂质。离心和过滤减少了过氧化氢处理的 CPS 的蛋白质污染物，以满足 WHO 规定的限制。通过进一步的核酸内切酶处理步骤除去剩余的核酸杂质，得到纯化的CPS。通过各种分析技术评估纯化 CPS 的表征，包括¹H NMR和竞争性抑制测定的抗原性。不同的过氧化氢浓度对 CPS 的抗原性有显着影响。然而，最佳工艺条件可以保留 CPS 的原生特性。