摘要
目的对C群脑膜炎球菌多糖(group C meningococcal polysaccharide,GCMP)蛋白结合物的5种制备方法进行比较。方法以GCMP作为多糖抗原,破伤风类毒素(tetanus toxoid,TT)和重组绿脓杆菌外毒素A(recombinant exoprotein A from Pseudomonas aeruginosa,r EPA)作为载体蛋白,采用5种结合路线制备7种结合物,并对其生化指标、免疫原性及磷酸铝佐剂的免疫增强效果进行检测。结果不同的结合方法均可将不同载体蛋白与多糖有效结合,在小鼠体内均具有良好的免疫原性。7种结合物的分子大小分布、游离多糖含量、多糖/蛋白值、回收率及免疫原性等指标有较大差异;采用相同结合方法,以TT和r EPA作为载体蛋白制备的结合物的生化指标及免疫原性无差异;磷酸铝佐剂可显著提高结合物的免疫原性。结论在选择GCMP蛋白结合物制备方法时应综合考虑原料多糖的化学结构、载体蛋白类型、结合路线、结合物纯化方法、质量控制方法和剂型等因素。本实验为GCMP蛋白结合物制备路线的选择提供了实验依据。
Objective To compare the five methods for preparation of group C meningococcal polysaccharide（GCMP）-protein conjugates. Methods Seven GCMP-protein conjugates were prepared by five binding approaches using GCMP as a polysaccharide antigen while tetanus toxoid（TT） and recombinant exoprotein A from Pseudomonas aeruginosa（r EPA）as carrier proteins, and analyzed for biochemical indexes, immunogenicity as well as enhancement of immune effect by aluminum phosphate adjuvant. Results Various carrier proteins and polysaccharides were bound effectively by various methods, which showed good immunogenicity in mice. Significant differences were observed in the molecular size distribution, free polysaccharide content, polysaccharide / protein ratio, recovery rate and immunogenicity of seven conjugates. No significant differences were observed in biochemical indexes or immunogenicity of conjugates prepared by the same methods using TT and r EPA as carrier proteins respectively. Conclusion the method for preparation of GCMPprotein conjugates should be selected based on the chemical structure of polysaccharide as material, type of carrier protein, binding approach, methods for purification and quality control as well as form. This study provided an experimental basis for selection of method for preparation of GCMP-protein conjugates.
出处
《中国生物制品学杂志》
CAS
CSCD
2015年第9期961-969,共9页
Chinese Journal of Biologicals
基金
国家科技支撑计划课题(2008BAI66B01)
甘肃省科技重大专项(0801NKDA003)
甘肃省"十二五陇药产业发展"专项
关键词
C群脑膜炎球菌多糖蛋白结合物
破伤风类毒素
重组绿脓杆菌外毒素A
磷酸铝佐剂
免疫原性
Group C meningococeal polysaccharide-protein conjugates
Tetanus toxoid （TF）
Recombinant exoprotein Afrom Pseudomonas aeruginosa （rEPA）
Aluminum phosphate adjuvant
Immunogenicity
