甲型副伤寒多糖-破伤风类毒素结合疫苗的制备及安全性和免疫原性

目的制备甲型副伤寒多糖-破伤风类毒素结合疫苗,并检测其安全性与免疫原性。方法副甲O-SP溶液加入CDAP活化,己二酰肼(ADH)衍化,在碳二亚胺(EDAC)作用下与破伤风类毒素(TT)偶联,反应物经柱层析纯化,制得多糖蛋白结合物,经质量检测后,进行安全性及免疫原性试验。结果所制备的结合物多糖蛋白比稳定在0.6~0.8之间,CL-4B检测Kd≤0.2时的回收率大于75%,高相对分子质量结合物含量不低于90%。免疫双扩散试验显示结合物与副甲家兔超免血清和TT抗血清均产生可见沉淀线。动物安全试验均合格。结合物免疫小鼠后,其血清副甲LPS抗体滴度显著增长,阳转率达到100%。结论用CDAP活化多糖制备的甲型副伤寒-破伤风类毒素结合疫苗质量稳定,安全可靠,且具有良好的免疫原性。

Preparation, Safety and Immunogenicity of S. paratyphoid A-Tetanus Toxoid Conjugate Vaccine

Objective To prepare S.paratyphoid A-tetanus toxoid(TT) conjugate vaccine and study its safety and immunogenicity.Methods The polysaccharide O-SP of S.paratyphoid A was activated with CDAP,derived with ADH,then coupled to TT using EDAC as condensing agent and purified by column chromatography to prepare S.paratyphoid A-tetanus toxoid(TT) conjugate.After control tests,the conjugate was further tested for safety and immunogenicity in animals.Results The ratio of polysaccharide to protein of the prepared conjugate was as stable as 0.6-0.8.The recovery rate of conjugate,determined by Sepharose Cl-4B chromatography,was more than 75% when Kd value was not more than 0.2.The content of high molecular conjugate was not less than 90%.Double immunodiffusion test showed visual precipitation lines of the conjugate with hyperimmune rabbit serum against S.paratyphoid A and TT antiserum.The conjugate showed good safety in animal test.The antibody titer against S.paratyphoid A of mice immunized with the conjugate increased significantly,and the antibody positive conversion rate reached 100%.Conclusion The S.paratyphoid A-tetanus toxoid(TT) conjugate vaccine prepared by activating polysaccharide O-SP with CDAP showed good stability,safety and immunogenicity.