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肺炎链球菌荚膜多糖分子质量较大,在制备结合疫苗的过程会有诸多不便,如溶液黏度大、结合过程易过度交联形成凝胶、不易于过滤以及纯化,而荚膜多糖分子片段化是降低其分子质量的有效途径。文章选取了酸水解和超声降解两种方法分别处理9V型肺炎链球菌荚膜多糖。利用不同相对分子质量的葡聚糖在HPLC上的保留时间绘制标准曲线,计算处理后多糖相对分子质量,并通过核磁共振检测结构稳定性,免疫双扩散检测抗原性。实验结果表明,在适宜的条件下,酸水解和超声降解均能有效降低肺炎链球菌荚膜多糖分子质量,减少重复单元,保留重复单元内部结构,保留时间从5.7 min延长到8.25 min,相对分子质量约为580 k。但是酸水解方法破坏了多糖的抗原完整性,而超声降解可以良好保持。
Streptococcus pneumoniae capsular polysaccharide molecular mass, in the process of preparation of conjugate vaccine will have a lot of inconvenience, such as solution viscosity, the process of crosslinking is too easy to form a gel, not easy to filter and purification, and capsular polysaccharide molecular fragmentation It is an effective way to reduce the molecular weight. In this paper, two methods of acid hydrolysis and ultrasonic degradation were used to treat 9V Streptococcus pneumoniae capsular polysaccharide. The retention time of HPLC-based dextran with different relative molecular mass was used to draw a standard curve, and the relative molecular mass of polysaccharides after treatment was calculated. The structural stability was detected by NMR and the antigenicity was detected by double immunodiffusion. The experimental results showed that acid hydrolysis and ultrasonic degradation could effectively reduce the molecular weight of S. pneumoniae capsular polysaccharides, reduce the number of repeat units and the internal structure of the repeat units, and the retention time extended from 5.7 to 8.25 min under appropriate conditions. The relative molecular weight The mass is about 580 k. However, acid hydrolysis destroys the antigenic integrity of the polysaccharide, and sonication can be well maintained.