{"title":"了解单价和五价（A、C、Y、W和X）脑膜炎球菌X非偶联糖的行为，用于脑膜炎球菌结合疫苗的质量分析。","authors":"Swapnil Phugare, Abhijit Patil, Sameer Kale, Pankaj Sharma, Sunil Kumar Goel, Sunil Gairola","doi":"10.1016/j.carres.2025.109678","DOIUrl":null,"url":null,"abstract":"<p><p>The pentavalent meningococcal polysaccharide conjugate vaccine, which includes serogroup A, C, Y, W, and X, has recently been prequalified by the WHO and is currently the only vaccine available against serogroup X. The amount of free saccharide in conjugate vaccines is a crucial factor that directly impacts the stability and immunogenicity of the vaccine. Therefore, precise estimation of free saccharide content is particularly important in multivalent conjugate vaccines. This study focuses on the development of conditions for estimating free saccharide in monovalent meningococcal X conjugate bulk. Accuracies were demonstrated at 5 %, 10 %, 25 %, and 40 % of the test specification, with recoveries ranging from 70 to 130 %. Repeatability analysis showed intra-assay variation ranging from 2 to 6 %, while inter-assay variation ranged from 2 to 14 %. Specificity studies indicated that there was no interference from assay components such as sample excipients, DOC, or acids. When these established conditions were applied to the finished product for free saccharide estimation, an interesting observation was noted. It was found that the unconjugated meningococcal X saccharide interacted with other biomolecules present in the vaccine formulation, leading to decreased free saccharide recovery for serogroup X. It is believed that aggregation, possibly due to carrier proteins (TT and CRM) and/or associated polysaccharides, may be responsible for trapping the free saccharide from serogroup X. Efforts were made to control these biomolecular interactions by adjusting solubilities, buffering, and physico-chemical conditions to separate free saccharide. Phosphate buffers, ionic-nonionic detergent solutions, salt buffers, and 32 biomolecule formulations were explored in various combinations before identifying the root cause for the decreased X free saccharide recoveries.</p>","PeriodicalId":9415,"journal":{"name":"Carbohydrate Research","volume":"558 ","pages":"109678"},"PeriodicalIF":2.5000,"publicationDate":"2025-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Understanding meningococcal X unconjugated saccharide behaviour in monovalent and pentavalent (A, C, Y, W and X) conjugate formulations for quality analysis of meningococcal conjugate vaccine.\",\"authors\":\"Swapnil Phugare, Abhijit Patil, Sameer Kale, Pankaj Sharma, Sunil Kumar Goel, Sunil Gairola\",\"doi\":\"10.1016/j.carres.2025.109678\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The pentavalent meningococcal polysaccharide conjugate vaccine, which includes serogroup A, C, Y, W, and X, has recently been prequalified by the WHO and is currently the only vaccine available against serogroup X. The amount of free saccharide in conjugate vaccines is a crucial factor that directly impacts the stability and immunogenicity of the vaccine. Therefore, precise estimation of free saccharide content is particularly important in multivalent conjugate vaccines. This study focuses on the development of conditions for estimating free saccharide in monovalent meningococcal X conjugate bulk. Accuracies were demonstrated at 5 %, 10 %, 25 %, and 40 % of the test specification, with recoveries ranging from 70 to 130 %. Repeatability analysis showed intra-assay variation ranging from 2 to 6 %, while inter-assay variation ranged from 2 to 14 %. Specificity studies indicated that there was no interference from assay components such as sample excipients, DOC, or acids. When these established conditions were applied to the finished product for free saccharide estimation, an interesting observation was noted. It was found that the unconjugated meningococcal X saccharide interacted with other biomolecules present in the vaccine formulation, leading to decreased free saccharide recovery for serogroup X. It is believed that aggregation, possibly due to carrier proteins (TT and CRM) and/or associated polysaccharides, may be responsible for trapping the free saccharide from serogroup X. Efforts were made to control these biomolecular interactions by adjusting solubilities, buffering, and physico-chemical conditions to separate free saccharide. Phosphate buffers, ionic-nonionic detergent solutions, salt buffers, and 32 biomolecule formulations were explored in various combinations before identifying the root cause for the decreased X free saccharide recoveries.</p>\",\"PeriodicalId\":9415,\"journal\":{\"name\":\"Carbohydrate Research\",\"volume\":\"558 \",\"pages\":\"109678\"},\"PeriodicalIF\":2.5000,\"publicationDate\":\"2025-09-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Carbohydrate Research\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://doi.org/10.1016/j.carres.2025.109678\",\"RegionNum\":3,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Carbohydrate Research","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1016/j.carres.2025.109678","RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Understanding meningococcal X unconjugated saccharide behaviour in monovalent and pentavalent (A, C, Y, W and X) conjugate formulations for quality analysis of meningococcal conjugate vaccine.
The pentavalent meningococcal polysaccharide conjugate vaccine, which includes serogroup A, C, Y, W, and X, has recently been prequalified by the WHO and is currently the only vaccine available against serogroup X. The amount of free saccharide in conjugate vaccines is a crucial factor that directly impacts the stability and immunogenicity of the vaccine. Therefore, precise estimation of free saccharide content is particularly important in multivalent conjugate vaccines. This study focuses on the development of conditions for estimating free saccharide in monovalent meningococcal X conjugate bulk. Accuracies were demonstrated at 5 %, 10 %, 25 %, and 40 % of the test specification, with recoveries ranging from 70 to 130 %. Repeatability analysis showed intra-assay variation ranging from 2 to 6 %, while inter-assay variation ranged from 2 to 14 %. Specificity studies indicated that there was no interference from assay components such as sample excipients, DOC, or acids. When these established conditions were applied to the finished product for free saccharide estimation, an interesting observation was noted. It was found that the unconjugated meningococcal X saccharide interacted with other biomolecules present in the vaccine formulation, leading to decreased free saccharide recovery for serogroup X. It is believed that aggregation, possibly due to carrier proteins (TT and CRM) and/or associated polysaccharides, may be responsible for trapping the free saccharide from serogroup X. Efforts were made to control these biomolecular interactions by adjusting solubilities, buffering, and physico-chemical conditions to separate free saccharide. Phosphate buffers, ionic-nonionic detergent solutions, salt buffers, and 32 biomolecule formulations were explored in various combinations before identifying the root cause for the decreased X free saccharide recoveries.
期刊介绍：
Carbohydrate Research publishes reports of original research in the following areas of carbohydrate science: action of enzymes, analytical chemistry, biochemistry (biosynthesis, degradation, structural and functional biochemistry, conformation, molecular recognition, enzyme mechanisms, carbohydrate-processing enzymes, including glycosidases and glycosyltransferases), chemical synthesis, isolation of natural products, physicochemical studies, reactions and their mechanisms, the study of structures and stereochemistry, and technological aspects.
Papers on polysaccharides should have a "molecular" component; that is a paper on new or modified polysaccharides should include structural information and characterization in addition to the usual studies of rheological properties and the like. A paper on a new, naturally occurring polysaccharide should include structural information, defining monosaccharide components and linkage sequence.
Papers devoted wholly or partly to X-ray crystallographic studies, or to computational aspects (molecular mechanics or molecular orbital calculations, simulations via molecular dynamics), will be considered if they meet certain criteria. For computational papers the requirements are that the methods used be specified in sufficient detail to permit replication of the results, and that the conclusions be shown to have relevance to experimental observations - the authors'' own data or data from the literature. Specific directions for the presentation of X-ray data are given below under Results and "discussion".
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