摘要
目的用不同活化剂进行22F型肺炎球菌荚膜多糖(type 22F streptococcus pneumoniae capsular polysaccharide)的活化,并探讨其多糖蛋白结合物的免疫原性。方法分别用溴化氰(cyanogen bromide,CNBr)和1-氰基-4-二甲氨基吡啶四氟硼酸酯(1-cyano-4-dimethylaminopyridinium tetrafluoroborate,CDAP)作为22F型肺炎球菌荚膜多糖的活化剂,1,6-己二酰肼(1,6-adipic acid dihydrazide,ADH)作为链接剂,制备22F型肺炎荚膜多糖衍生物(3批Pn22FpsADH1和3批Pn22Fps-ADH2);在碳二亚胺[N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride,EDAC]的作用下,将衍生物与破伤风类毒素(tetanus toxoid,TT)共价结合,经凝胶过滤柱层析纯化,得到22F型肺炎链球菌荚膜多糖蛋白结合物(3批Pn22Fps-TT1和3批Pn22Fps-TT2);并对其生化指标、血清学特异性、免疫原性及异常毒性进行检测。结果衍生物Pn22Fps-ADH2的衍生率及回收率均略高于Pn22Fps-ADH1;结合物Pn22Fps-TT2的生化指标检测结果相对Pn22Fps-TT1较好;结合物Pn22Fps-TT1和Pn22Fps-TT2均可与22F型肺炎球菌诊断血清特异性结合;结合物Pn22Fps-TT1和Pn22Fps-TT2均具有良好的免疫原性;无异常毒性。结论 CNBr和CDAP均可作为22F型肺炎球菌荚膜多糖活化剂,经活化和结合反应,其抗原位点得到较好保留,但CDAP作为结合疫苗多糖的活化试剂效果更佳。
Objective To activate type 22 F Streptococcus pneumoniae capsular polysaccharide with different activators and investigate the immunogenicity of polysaccharide- protein conjugate. Methods Using cyanogen bromide(CNBr)and1-cyano-4-dimethylamino pyridinium tetrafluoroborate(CDAP) as activators and 1, 6- adipic acid dihydrazide(ADH) as a linker, type 22 F pneumoniae capsular polysaccharide derivatives(Pn22Fps-ADH)were prepared, including three batches of Pn22Fps-ADH1 and three batches of Pn22Fps-ADH2. The derivatives were covalently conjugated to tetanus toxoid(TT)using N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride(EDAC)as a coupling agent, then purified by gel filtration column chromatography to obtain type 22 F S. pneumoniae capsular polysaccharide-protein conjugates(Pn22Fps-TT), including three batches of Pn22Fps-TT1 and three batches of Pn22Fps-TT2. The biochemical indexes,serological specificity, immunogenicity and abnormal toxicity of the conjugates were determined. Results Both the derivation and recovery rates of Pn22Fps-ADH2 were slightly higher than those of Pn22Fps-ADH1. The biochemical indexes of Pn22Fps-TT2 were superior to those of Pn22Fps-TT1. Both Pn22Fps-TT1 and Pn22Fps-TT2 showed specific binding to diagnostic sera against type 22 F S. pneumoniae as well as high immunogenicity, while showed no abnormal toxicity.Conclusion Both CNBr and CDAP may be used as the activators of type 22 F pneumococcal capsular polysaccharide.The antigenic epitope of type 22 F pneumococcal capsular polysaccharide was retained well through activation and binding.However, CDAP was a more effective activator of polysaccharide of conjugate vaccine.
出处
《中国生物制品学杂志》
CAS
CSCD
2015年第5期479-482,487,共5页
Chinese Journal of Biologicals
关键词
肺炎链球菌荚膜多糖
溴化氰
1-氰基-4-二甲氨基吡啶四氟硼酸酯
破伤风类毒素
免疫原性
Streptococcus pneumoniae capsular polysaccharide Cyanogen bromide(CNBr) 1-Cyano-4-dimethylamino pyridinium tetrafluoroborate(CDAP) Tetanus toxoid(TT) Immunogenicity