核磁共振波谱法在肺炎球菌荚膜多糖检定中的应用

Application of NMR spectrum in identification of pneumococcal capsular polysaccharides

目的建立1H-NMR波谱法检定23价肺炎球菌荚膜多糖结构的方法。方法用1H-NMR方法分析肺炎球菌荚膜多糖,选择(5.89～4.64)ppm区作为‘指纹’鉴定区,以默克公司公布的肺炎球菌荚膜多糖波谱为参照谱图,将检定样品的波谱与其相比较。对1H-NMR波谱进行专属性和耐用性验证。结果实验中应用1H-NMR波谱分析法对23价肺炎球菌多糖疫苗包含的所有血清型多糖的检定结果与默克公司公布的结果完全一致,该方法有很好的专属性和耐用性。结论实验中建立的1H-NMR波谱法适用于肺炎球菌荚膜多糖的检定,与传统方法相比,该方法有检测速度快,样品易于准备的优点等。

Objective To establish a NMR-based identity assay for pneumococcal capsular polysaccharides. Methods Pneumococcal capsular polysaccharides were identified with 1H-NMR spectrum. A portion of the anomeric region of each spectrum （5.89 to 4.64 ppm} was selected as the ＇fingerprint＇ identification region. The spetrum of tested samples were compared with the reference spectrum which were published by Merck Co. , Ltd. Results When 1H-NMR was used in analysis of pneumococcal capsular polysaccharides samples, there was no difference in between the tested sample spectrum and the reference spectrum. The specificity and robustness of 1H-NMR-based identity assay is superior to conventional col- orimetric method. Conclusions The NMR method established in our laboratory is suitable for PnPs identification. The NMR assay is more easy to operate and time saving than conventional methods.