| 引用本文: | 郑佳睿,秦春君,胡静,任涛,尹珊珊,刘建凯,尹健.肺炎链球菌荚膜多糖氰酸酯化中生成基团测定方法的适用性研究[J].中国现代应用药学,2025,42(11):99-105. |
| ZHENG Jiarui,QIN Chunjun,HU Jing,REN Tao,YIN Shanshan,LIU Jiankai,YIN Jian.Applicability of Quantitative Determination Methods for Groups Generated in Cyanate Esterification of Pneumococcal Capsular Polysaccharides[J].Chin J Mod Appl Pharm(中国现代应用药学),2025,42(11):99-105. |
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| 摘要: |
| 目的 针对已报道可定量测定氰酸酯、亚胺碳酸酯、氨基甲酸酯三种基团的方法，探究其对肺炎链球菌荚膜多糖氰酸酯化中各生成基团的适用性。方法 对于活化多糖氰酸酯基含量，使用高氯酸滴定法和二甲基巴比妥酸吡啶法进行测定；对于亚胺碳酸酯基含量的测定，使用盐酸水解法将亚胺碳酸酯水解为铵离子，通过分光光度法测得亚胺碳酸酯基含量；对于氨基甲酸酯基含量，使用酶联免疫法进行测定。结果 由于肺炎链球菌荚膜多糖在乙酸中溶解性较差，高氯酸滴定法对于该类多糖活化物中氰酸酯基含量测定有待进行反应溶剂的优化。通过二甲基巴比妥酸吡啶法测定23F型肺炎链球菌荚膜多糖活化后的氰酸酯基生成率为0.038%±0.004%。盐酸水解法测定23F型肺炎链球菌荚膜多糖活化后的亚胺碳酸酯基生成率为2.20%±0.03%。酶联免疫法测定的活化多糖氨基甲酸酯基含量结果显示超过检测范围，该方法不适用于活化多糖的反应体系。结论 二甲基巴比妥酸吡啶法可测定活化程度达到0.038%±0.004%的氰酸酯化多糖中氰酸酯基含量。盐酸水解法可以准确测定氰酸酯化多糖的亚胺碳酸酯基含量。酶联免疫法并不适用于氰酸酯化多糖的氨基甲酸酯基含量测定。 |
| 关键词: 肺炎链球菌 荚膜多糖 多糖结合疫苗 多糖氰酸酯化 亚胺碳酸酯基 氨基甲酸酯基 |
| DOI： |
| 分类号:R917 |
| 基金项目:国家自然科学基金(22325803，22277042，22077052，22177041) |
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| Applicability of Quantitative Determination Methods for Groups Generated in Cyanate Esterification of Pneumococcal Capsular Polysaccharides |
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ZHENG Jiarui1, QIN Chunjun1, HU Jing2, REN Tao3, YIN Shanshan3, LIU Jiankai3, YIN Jian1,4
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1.Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University;2.Wuxi School of Medicine, Jiangnan University;3.Beijing Engineering Technology Research Center for New Combined Vaccine, Beijing Minhai Biotechnology Co., Ltd.;4.School of Life Sciences and Health Engineering, Jiangnan University
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| Abstract: |
| OBJECTIVE Applicabilities of previously reported quantitative determination methods for cyanate ester, imidocarbonate, and carbamate groups for CDAP-activated pneumococcal capsular polysaccharides (CPS) were explored. METHODS For the content of cyanate ester in activated polysaccharide, the perchloric acid titration method and the dimethyl barbiturate pyridine method were used. For the determination of imidocarbonate content, the hydrochloric acid hydrolysis method was used to hydrolyze imine carbonate into ammonium ions, which was measured by spectrophotometry. For the content of carbamate, enzyme-linked immunosorbent assay was used. RESULTS Due to the insolubility of the Streptococcus pneumoniae CPS in acetic acid, the determination of cyanate ester content in the activated S. pneumoniae CPS by using perchloric acid titration requires optimization of the reaction solvent. On the other hand, the cyanate ester content of activated 23F S. pneumoniae CPS determined by the dimethylbarbiturate pyridine method was 0.038%±0.004%. The imidocarbonate content of 23F S. pneumoniae CPS determined by hydrochloric acid hydrolysis method was 2.20%±0.03%. The aminoformate content of activated polysaccharide determined by enzyme-linked immunosorbent assay exceeded the detection range. This method was thought to be not suitable for the CDAP-activated pneumococcal polysaccharides. CONCLUSION The dimethyl barbiturate pyridine method can be used to determine the cyanate ester content in cyanated polysaccharides with an activation degree of 0.038%±0.004%. The hydrochloric acid hydrolysis method can accurately determine the imidocarbonate content of CDAP-activated polysaccharides. Enzyme linked immunosorbent assay was found to be not suitable for the determination of carbamate content in CDAP-activated polysaccharides. |
| Key words: Streptococcus pneumoniae capsular polysaccharides glycoconjugate vaccine cyanate esterification of polysaccharide imidocarbonate carbamate |
