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
郭吉茹 1秦春君 1胡静 2曹欣 3徐永学 3刘建凯 3尹健4
作者信息
- 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. 江南大学生物工程学院糖化学与生物技术教育部重点实验室,无锡 214122;江南大学生命科学与健康工程学院,无锡 214122
- 折叠
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.
Abstract
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Key words
Streptococcus pneumoniae/capsular polysaccharide/quantitative 1H NMR/quantitative 31P NMR/C-polysaccharide/phosphorus引用本文复制引用
基金项目
国家自然科学基金(22325803)
国家自然科学基金(22277042)
国家自然科学基金(22077052)
国家自然科学基金(22177041)
出版年
2024
国家科技期刊平台
NSTL
万方数据