论文部分内容阅读
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 22F streptococcus pneumoniae capsular polysaccharide with different activators and investigate the immunogenicity of its polysaccharide conjugate. Methods Cyanogen bromide (CNBr) and 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) were used as capsular polysaccharide of pneumococcal 22F 22F pneumococcal capsular polysaccharide derivatives (3 batches of Pn22FpsADH1 and 3 batches of Pn22Fps-ADH2) were prepared using linoleic acid, 1,6-adipic acid dihydrazide (ADH) Derivatives were covalently bound to tetanus toxoid (TT) under the action of N-ethylcarbodiimide hydrochloride (N- (3-Dimethylaminopropyl) -N’-ethylcarbodiimide hydrochloride, purified by gel filtration chromatography, The 22F pneumococcal capsular polysaccharide conjugates (3 batches of Pn22Fps-TT1 and 3 batches of Pn22Fps-TT2) were obtained. The biochemical indexes, serological specificity, immunogenicity and abnormal toxicity were also detected. Results Pn22Fps-ADH2 derivative and recovery rates were slightly higher than Pn22Fps-ADH1; Pn22Fps-TT2 conjugate Pn22Fps-TT1 biochemical detection results are better; conjugates Pn22Fps-TT1 and Pn22Fps-TT2 with 22F Pneumococcal diagnosis of serum-specific binding; conjugates Pn22Fps-TT1 and Pn22Fps-TT2 have good immunogenicity; no abnormal toxicity. CONCLUSION Both CNBr and CDAP can be used as 22F pneumococcal capsular polysaccharide activator. The activation and binding of CNBr and CDAP keep their antigenic sites well. However, CDAP is more effective as an activator of polysaccharide conjugate vaccine.