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目的研究人乳头瘤病毒（ＨＰＶ）相关肿瘤患者的ＨＰＶ１６Ｅ６抗体水平及其流行病学意义；探讨用杆状病毒－昆虫细胞表达载体系统表达的ＨＰＶ１６Ｅ６蛋白的抗原性。方法用ＰＣＲ从ＨＰＶ１６基因组中扩增出ＨＰＶ１６Ｅ６完整基因，克隆至转移载体ｐＶＬ１３９３中，重组质粒ＤＮＡ与线性杆状病毒ＤＮＡ共转染昆虫细胞Ｓｆ－９，经噬斑筛选获得带有编码Ｅ６基因的重组杆状病毒株，并在昆虫细胞中表达。结果Ｗｅｓｔｅｒｎｂｌｏｔ和高效液相色谱法检测ＨＰＶ１６Ｅ６表达蛋白，其分子量约为１８０００。免疫印迹检测显示其能与兔抗ＨＰＶ１６Ｅ６多抗特异性结合。酶联免疫吸附试验表明此重组蛋白能被人ＨＰＶ１６阳性血清所识别。结论昆虫细胞表达的ＨＰＶ１６Ｅ６蛋白，具有良好的抗原性，可用于检测ＨＰＶ１６Ｅ６特异性免疫球蛋白ＩｇＧ和ＩｇＭ抗体
Objective To study the level of HPV16E6 antibody in patients with human papillomavirus (HPV) -related tumors and its epidemiological significance. To explore the antigenicity of HPV16E6 protein expressed in baculovirus-insect cell expression vector system. Methods The full-length HPV16E6 gene was amplified by PCR from HPV16 genome and cloned into pVL1393. The recombinant plasmid DNA was co-transfected with Sf-9 insect baculovirus DNA. Baculovirus strains are recombinant and expressed in insect cells. Results HPV16E6 protein was detected by Western blot and HPLC. The molecular weight of HPV16E6 protein was about 18000. Western blotting showed that it could specifically bind to rabbit anti-HPV16E6 polyclonal antibody. Enzyme-linked immunosorbent assay showed that the recombinant protein was recognized by human HPV16 positive serum. Conclusion The HPV16E6 protein expressed in insect cells has good antigenicity and can be used to detect HPV16E6 specific immunoglobulin IgG and IgM antibodies