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为在真核细胞中表达并纯化I型单纯疱疹病毒(HSV I)包膜糖蛋白gB,并分析其抗原性和免疫原性,化学合成了包膜糖蛋白gB1胞外区基因片段,构建真核表达载体,并转染至HEK293细胞,表达的蛋白用羊抗HSV1+HSV2血清作为一抗,用ELISA检测其抗原性;用纯化的gB1蛋白免疫昆明小鼠,观察诱发抗体产生的时间及其效价,并用ELISA和Western blot检测小鼠抗gB1多克隆抗体特异性识别重组gB1抗原的能力,评价其免疫原性。结果显示在HEK293细胞中成功表达重组gB1蛋白,ELISA证实羊抗HSV1+HSV2多抗能够识别重组gB1蛋白;重组gB1蛋白免疫小鼠7周后,小鼠血清中多克隆抗体效价达到5×103,表明在真核细胞中高效表达并纯化的重组gB1蛋白具有良好的抗原性和免疫原性,为HSV检测试剂和疫苗研究提供了理论基础。
In order to express and purify HSV I envelope glycoprotein gB in eukaryotic cells and analyze its antigenicity and immunogenicity, the envelope glycoprotein gB1 extracellular region gene was chemically synthesized and constructed And then transfected into HEK293 cells. The expressed protein was detected by ELISA using goat anti-HSV1 + HSV2 serum as the primary antibody. The purified gB1 protein was used to immunize Kunming mice to observe the time of induced antibody production and its The anti-gB1 polyclonal antibody of mouse was used to detect the specificity of recombinant gB1 antigen by ELISA and Western blot, and its immunogenicity was evaluated. The results showed that recombinant gB1 protein was successfully expressed in HEK293 cells. ELISA confirmed that goat anti-HSV1 + HSV2 polyclonal antibody could recognize recombinant gB1 protein. After 7 weeks of recombinant gB1 protein immunization, the titer of polyclonal antibody in mouse serum reached 5 × 103 , Indicating that the recombinant gB1 protein highly expressed and purified in eukaryotic cells has good antigenicity and immunogenicity and provides a theoretical basis for the research on HSV detection reagents and vaccines.