Department of Technology, Savitribai Phule Pune University, Pune 411007, India; Research and Development Department, Serum Institute of India Pvt. Ltd., Hadapsar, Pune 411028, Maharashtra, India.
Research and Development Department, Serum Institute of India Pvt. Ltd., Hadapsar, Pune 411028, Maharashtra, India.
Carbohydr Polym. 2022 Oct 15;294:119783. doi: 10.1016/j.carbpol.2022.119783. Epub 2022 Jun 27.
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 H 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.
对于生产糖缀合物疫苗来说，高质量且具有成本效益的纯化程序是最重要的要求之一。本工作的目的是设计一种从肺炎链球菌血清型 2 荚膜多糖（CPS）中去除杂质（如蛋白质和核酸）的方法。研究了使用过氧化氢来降低粗 CPS 中的杂质。过氧化氢处理后的 CPS 经离心和过滤可降低蛋白质污染物含量，达到世卫组织规定的限度。通过进一步的内切核酸酶处理去除残留的核酸杂质，得到纯化的 CPS。通过各种分析技术（包括 1H NMR 和竞争性抑制测定法的抗原性）对纯化的 CPS 进行了表征。不同浓度的过氧化氢对 CPS 的抗原性有显著影响。然而，最佳的工艺条件可以保持 CPS 的天然特性。
