【作者】 刘方蕾； 吴兵； 王晶； 袁菲； 赵志强； 杨明； 谢贵林；

【Author】 LIU Fang-lei,WU Bing,WANG Jing,YUAN Fei,ZHAO Zhi-qiang,YANG Ming,XIE Gui-lin (Lanzhou Institute of Biological Products,Lanzhou 730046,China)

【机构】 兰州生物制品研究所第一研究室；

【摘要】 目的:按初步确定的工艺,制备多批A群奈瑟氏脑膜炎球菌荚膜多糖(GAMP)与破伤风类毒素(TT)的结合物并检测其免疫原性,通过监测关键工艺参数,对该工艺的稳定性进行研究。方法:采用溴化氰(CNBr)活化多糖,以无水己二酸二肼(ADH)作为连接剂,1-乙基1-3-(3-二甲基氨基-丙基)-碳化二亚胺(EDAC)为偶联剂制备8批结合物;以结合物皮下免疫小鼠,ELISA检测小鼠血清中抗GAVMP及抗载体蛋白IgG抗体水平。结果:艺过程中关键参数较稳定。制备的各批多糖-蛋白质结合物都保持GAMP抗原特异性;结合物免疫小鼠后均可诱生较高水平的血清抗GAMP IgG抗体和免疫加强应答。结论:该制备工艺稳定,可为A群奈瑟氏脑膜炎球菌荚膜多糖(GAMP)与破伤风类毒素(TT)结合疫苗的研制提供有力依据。更多还原

【Abstract】 Objective:Different lots of GAMP-TT conjugate are prepared according to the determinate technology.Key technical parameters and the immunogenicity of conjugates are investigated to evaluate the stability of technology. Methods:After the polysaccharide was activated by cyanogen Bromide(CNBr),8 lots of GAMP-TT conjugates were prepared using adipic Aciddihydrazide(ADH) as a linker and l-ethyl-3-(3-dimethylamino-propyl) Carbodiimide(EDAC) as coupling reagent.Mice were subcutaneously immunized with the conjugates and serum IgG antibodies against GAMP and TT were measured by enzyme-linked immunosorbent assay(ELISA).Results:Technical parameters are consistent among different lots of conjugates.All conjugates maintain the specificity of GAMP.High levels of serum antibodies to GAMP and booster response were induced in immunized mice.Conclusion:The technological stability for preparation of GAMP-TT conjugate were proved,which will facilitate the development of GAMP-TT conjugate vaccine.更多还原