CDAP活化多糖制备5型肺炎链球菌荚膜多糖-破伤风类毒素结合疫苗

Preparation of Type 5 Streptococcus pneumoniae Capsular Polysaccharide-TT Conjugate Vaccine by Activation of Polysaccharide with CDAP

目的用1-氰基-4-二甲氨基-吡啶四氟硼酸(CDAP)代替溴化氰(CNBr)活化5型肺炎链球菌荚膜多糖,制备5型肺炎链球菌荚膜多糖-破伤风类毒素(TT)结合疫苗。方法将5型肺炎链球菌荚膜多糖溶液加CDAP活化,经ADH和缩合剂EDAC与破伤风类毒素进行偶联,经凝胶过滤柱层析纯化,得到5型肺炎链球菌多糖-TT结合物,并对其生化指标、血清学特异性、安全性及免疫原性进行检测。结果多糖-TT结合物的游离多糖含量与多糖的衍化率成反比;血清学特异性良好;经动物实验证明其安全性合格;具有良好的免疫原性,且游离多糖含量越低,免疫原性越强。结论用CDAP活化工艺制备的5型肺炎链球菌荚膜多糖-TT结合物适宜制备疫苗。

Objective To prepare type 5 Streptococcus pneumoniae capsular polysaccharide-TT conjugate vaccine by activation of polysaccharide with 1-cyano-4-dinmethylaminopyridinium tetrafluoroborate（CDAP） instead of cyanogen bromide（CNBr）.Methods Type 5 Streptococcus pneumoniae capsular polysaccharide solution was activated with CDAP and coupled to TT by using ADH and condensing agent EDAC,then purified by gel filtration column chromatography to prepare type 5 Streptococcus pneumoniae capsular polysaccharide-TT conjugate which was determined for biochemical indexes and evaluated for serological specificity,safety and immunogenicity.Results The free polysaccharide content in the prepared conjugate was negatively related to the derivation rate of polysaccharide.The conjugate showed high specificity and immunogenicity as well as safety in animal test,and its immunogenicity increased with the decreasing free polysaccharide content.Conclusion The type 5 Streptococcus pneumoniae capsular polysaccharide-TT conjugate prepared by activation with CDAP is suitable for development of vaccine.