单一内标定量核磁法测定肺炎链球菌多糖抗原中荚膜多糖、C-多糖和磷含量  

Quantitative determination of capsular polysaccharide,C-polysaccharide,phosphorus of carbohydrate antigens from Streptococcus pneumoniae by quantitative NMR using a single internal standard

作　　者：郭吉茹 秦春君 胡静 曹欣[3] 徐永学 刘建凯[3] 尹健 GUO Jiru;QIN Chunjun;HU Jing;CAO Xin;XU Yongxue;LIU Jiankai;YIN Jian(Key Laboratory of Carbohydrate Chemistry and Biotechnology(Ministry of Education),School of Biotechnology,Jiangnan University,Wuxi 214122;Wuxi School of Medicine,Jiangnan University,Wuxi 214122;Beijing Key Laboratory of New Technology Research of Conjugate Vaccine,Beijing Minhai Biotechnology Co.,Ltd.,Beijing 102600;School of Life Sciences and Health Engineering,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学生物工程学院糖化学与生物技术教育部重点实验室,无锡214122 [2]江南大学无锡医学院,无锡214122 [3]北京民海生物科技有限公司结合疫苗新技术研究北京市重点实验室,北京102600 [4]江南大学生命科学与健康工程学院,无锡214122

出　　处：《中国药科大学学报》2024年第4期472-477,共6页Journal of China Pharmaceutical University

基　　金：国家自然科学基金项目(No.22325803,No.22277042,No.22077052,No.22177041)。

摘　　要：肺炎糖疫苗在预防由肺炎链球菌引起的肺炎和脑膜炎等疾病中发挥关键作用。荚膜多糖、C-多糖和磷含量是疫苗研发和生产中多糖抗原质量控制的重要指标,研究发展了基于单一内标物六甲基磷酰三胺(hexamethylphosphoramide,HMPA)的定量1H NMR和31P NMR法,实现6A,6B,18C,19A,19F及23F型肺炎链球菌多糖抗原中荚膜多糖、C-多糖和磷含量的同步测定。利用内参物比较法,探究了多糖溶解性质对定量核磁测定的影响,发现定量1H NMR的测定结果受到多糖水溶液黏度和浓度的影响。发现高黏度多糖在3~15 mg/mL,低黏度多糖在5-25 mg/mL是最适检测液浓度范围。该“一内标三定量”NMR法具有良好的精密度、特异性和准确度,为肺炎链球菌疫苗的质量控制提供了一种有价值的新策略。Pneumococcal vaccine plays a key role in preventing diseases such as pneumonia and meningitis caused by Streptococcus pneumoniae(S.pneumoniae).Capsular polysaccharide,C-polysaccharide and phosphorus content are important indicators for quality control of polysaccharide antigens in vaccine development and production.In this study,a quantitative 1H NMR and 31P NMR method based on a single internal standard hexamethylphosphoramide(HMPA)was developed to achieve simultaneous determination of capsular polysaccharide,C-polysaccharide and phosphorus content in 6A,6B,18C,19A,19F and 23F S.pneumoniae polysaccharide antigens.Using the internal reference comparison method,the effect of solubility of polysaccharide on quantitative 1H NMR determination was investigated.It was found that the determination results of quantitative 1H NMR were affected by the viscosity and concentration of polysaccharide solution.It was found that high viscosity polysaccharides at 3−15 mg/mL and low viscosity polysaccharides at 5−25 mg/mL were the optimal detection solution concentration range.This“one internal standard three quantitative”NMR method has good precision,specificity and accuracy,and provides a valuable new strategy for the quality control of pneumococcal vaccine.

关 键 词：肺炎链球菌 荚膜多糖 定量1H NMR 定量31P NMR C-多糖 磷 

分 类 号：R917[医药卫生—药物分析学]