A群脑膜炎球菌多糖—精破类偶联抗原的制备及其主要特性

Preparation, Physiochemical Characteristics and Stability of Meningococcal Group A Polysaccharide-Tetanus Toxoid Conjugate Antigen

参照生物制品质量要求,连续制备3批脑膜炎球菌多糖-精破类偶联抗原,其多糖含量为19.8～24.4%(w/w),该种拟糖蛋白具有波长为245nm特征性的紫外光吸收谱,在Sepharose-CL 4B柱层析上Kd为0。偶联抗原中多糖的抗原性在火箭电泳上虽与单纯多糖相似,但其免疫原性则明显增强。偶联抗原在4℃存放3年或在37℃存放8周,其免疫原性无明显改变。加温处理过的偶联抗原经Sepharose-CL4B和SDSPAGE检查表明,多糖与蛋白共价结合是相当稳定的。

According to quality requirement of biologics, we prepared three successive lots of Meningococcal Group A Polysaccharide-Tetanus Toxcid Conjugate Vaccine, which was composed of 19.8-24.4% (w/w) Polysaccharide moiety. The neoglycoprotein had a characteristic ultraviolet spectrum of 245nm and Kd value of 0 on Sepharose-CL4B.The immunogenicity of Ps moiety of the conjugate was markedly enhanced although the antigenicity of Ps moiety was the same as plain polysaccharide on the rocket immunoelectrophoresis.The stability of the conjugates was examined. There was no significant difference in immunogenicity of the conjugates during 3-year storage at 4℃ or 8-week storage at 37℃. The tests using gel permeation chromatography with Sepharose-CL4B and SDS-PAGE strongly affirmed the covalency of the derivatized protein-polysaccharide linkage and its stability through high temperature treatment.