【摘要】 为制备鼠疫耶尔森氏菌F1-V重组融合蛋白抗原,观察其免疫原性和免疫保护效果,通过疏水层析、阴离子交换层析、凝胶过滤层析纯化鼠疫F1-V重组融合蛋白抗原.用氢氧化铝凝胶吸附制备试验性鼠疫F1-V重组融合蛋白抗原,皮下接种健康BALB/c小鼠,ELISA检测血清F1-V抗体效价、MTT法测定淋巴细胞增殖能力,进一步用400LD50鼠疫耶尔森氏菌141标准毒株皮下攻毒,观察动物的存活情况.通过三步柱层析纯化获得的鼠疫F1-V重组融合蛋白抗原纯度达到90%以上.氢氧化铝凝胶吸附的鼠疫F1-V重组融合蛋白抗原免疫BALB/c小鼠三次,血清抗F1-V抗体效价为1∶(51200±800),对耶尔森氏菌141强毒株攻击的保护率是90%.上述结果表明,制备的鼠疫F1-V重组融合蛋白抗原具有良好的免疫原性和免疫保护效果,为研制鼠疫F1-V重组融合蛋白疫苗奠定了基础.更多还原

【Abstract】 F1-V recombinant fusion protein was purified to immunize mice and its immunogenicity and immuno-protection are detected. Different purification strategies were used to purify F1-V recombinant fusion protein. First hydrophobic chromatography, then anion exchange chromatography, and finally size exclusion chromatography. Aluminium hydroxide gel adsorbed F1-V fusion protein was intramuscularly injected into BALB/c mice. Anti-serum titer and protection ratio were detected. The purity of F1-V recombinant fusion protein was more than 90%. The anti-serum titer was high up to 1:(51 200 ?à 800). The protein antigen was capable of inducing an effective immuno-protection against subcutaneous challenge of 400 LD50 virulent Yersinia pestis with the survival rate of 90%. The F1-V recombinant fusion protein was able to protect against subcutaneous challenge of virulent Yersinia pestis, suggesting that the F1-V substance is well suited for developmentas the active ingredient of the next plague vaccine.更多还原