CAJ | 학술논문

目的建立Y群和W135群脑膜炎球菌多糖结合疫苗的制备方法.方法将脑膜炎球菌荚膜多糖以溴化氰活化及己二酰肼衍生制备多糖衍生物.对衍生条件进行优化,选取适宜衍生条件生产的衍生物与破伤风类毒素在碳二亚胺作用下结合,制成结合疫苗.将疫苗免疫小鼠,检测小鼠血清中抗Y群和W135群脑膜炎球菌多糖抗体滴度.用双向免疫扩散法对结合物进行抗原性检测.采用t检验对结果进行统计学分析.结果用15 g/L多糖在pH 8条件下活化时间10 min以及溴化氰/多糖质量比为1.5的衍生工艺制备Y群多糖衍生物,用15 g/L多糖在pH 7的条件下活化时间30 min以及溴化氰/多糖质量比为1.0的衍生工艺制备W135群多糖衍生物.所制备的结合物稳定性良好并且具有较好的免疫原性和抗原性.与多糖相比,多糖蛋白结合物可诱导较高水平的抗体应答,并且第2和第3针免疫后的抗体滴度明显高于第1和第2针(Y群:t=9.151、3.374,P＜0.01；W135群:t=17.21、2.44,P＜0.05).结论经过筛选优化的衍生工艺适合于多糖结合疫苗的研制,由此成功建立了Y群和W135群脑膜炎球菌多糖结合疫苗的制备方法.
목적 건립Y군화W135군뇌막염구균다당결합역묘적제비방법.방법 장뇌막염구균협막다당이추화청활화급기이선정연생제비다당연생물.대연생조건진행우화,선취괄의연생조건생산적연생물여파상풍류독소재탄이아알작용하결합,제성결합역묘.장역묘면역소서,검측소서혈청중항Y군화W135군뇌막염구균다당항체적도.용쌍향면역확산법대결합물진행항원성검측.채용t검험대결과진행통계학분석.결과 용15 g/L다당재pH 8조건하활화시간10 min이급추화청/다당질량비위1.5적연생공예제비Y군다당연생물,용15 g/L다당재pH 7적조건하활화시간30 min이급추화청/다당질량비위1.0적연생공예제비W135군다당연생물.소제비적결합물은정성량호병차구유교호적면역원성화항원성.여다당상비,다당단백결합물가유도교고수평적항체응답,병차제2화제3침면역후적항체적도명현고우제1화제2침(Y군:t=9.151、3.374,P＜0.01；W135군:t=17.21、2.44,P＜0.05).결론 경과사선우화적연생공예괄합우다당결합역묘적연제,유차성공건립료Y군화W135군뇌막염구균다당결합역묘적제비방법.
Objective To establish a method for preparation of groups Y and W135 meningococcal polysaccharide conjugate vaccines.Methods Groups Y (GYMP) and W135 (GWMP) meningococcal polysaccharides were activated with cyanogen bromide (CNBr) and linked with adipic acid dihydrazide.The derivation processes of polysaccharide were optimized and the derivatives produced in the suitable condition were conjugated to tetanus toxoid (TT).Mice were immunized with GYMP-TT and GWMP-TT,and determined for antibodies against GYMP and GWMP in sera.Antigenicity of the conjugates was detected by double immunodiffusion.The statistical analysis of results was made by t test.Results Group Y and group W135 polysaccharide derivatives were produced with 15 g/L GYMP activated at pH 8 for 10 min (CNBr/ GYMP =1.5) and 15 g/L GWMP activated at pH 7 for 30 min (CNBr/GWMP =1.0),respectively.Both group Y and group W135 meningococcal conjugate vaccines prepared with the optimized polysaccharide derivatives showed high stability,immunogenicity and antigenicity.The mice immunized with conjugate vaccines produced more antibodies than those with polysaccharides.Antibody levels induced with the second and third doses of conjugate vaccines were significantly higher than those with the first and second doses (group Y:t=9.151,3.374,P＜0.01; group W135:t=17.21,2.44,P＜0.05),respectively.Condusions The optimized derivation processes of polysaccharides are suitable for preparation of conjugate vaccines.The method for preparation of groups Y and W135 meningococcal polysaccharide conjugate vaccine is successfully developed.