神经药理学报

神经药理学报 ›› 2025, Vol. 15 ›› Issue (1): 1-.DOI: 10.3969/j.issn.2095-1396.2025.01.001

• 研究论文 •    下一篇

人乳头瘤病毒16 和18 型的E6 和E7 蛋白多克隆抗体制备及鉴定

周艳，张婷，王志荣，许雪梅   

  1. 1. 河北北方学院医学检验学院免疫教研室，张家口，075000，中国 

    2. 北京协和医学院基础医学研究所，北京，100010，中国

  • 出版日期:2025-02-26 发布日期:2025-06-20
  • 通讯作者: 许雪梅，教授，博士生导师
  • 基金资助:
    CAMS 医学科学创新基金项目(No. 2021-I2M-1-043)；河北北方学院省属高校基本科研业务费（No.JYT2022005）

Preparation and Identification of Polyclonal Antibodies Against E6 and E7 Proteins of Human Papillomavirus 16 and 18

ZHOU Yan, ZHANG Ting, WANG Zhi-rong, XU Xue-mei   

  1. 1. Department of Immunology, School of Laboratory Medicine, Hebei North University, Zhangjiakou, 075000, China 

    2. Institute of Basic Medicine, Peking Union Medical College, Beijing, 100010, China

  • Online:2025-02-26 Published:2025-06-20
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摘要：

目的：预测人乳头瘤病毒（human papillomavirus, HPV）16 型和18 型的E6 及 E7 蛋白的B 细胞线性表 位，制备兔抗HPV16 和HPV18 的E6 和E7 的多克隆抗体。方法：利用生物信息学软件分析HPV16 和HPV18 的E6 和E7 蛋白的氨基酸序列，预测B 细胞线性表位，设计并人工合成HPV16 E6、HPV16 E7、HPV18 E6 及 HPV18 E7 多肽，将合成肽分别与血蓝蛋白（keyhole limpet hemocyanin，KLH）偶联，免疫新西兰大白兔，制备 HPV16 和HPV18 的E6 和E7 多克隆抗体。以原核系统表达的重组蛋白Flic-HPV16 E6 E7、Flic-HPV18 E6 E7 为检测抗原，采用间接ELISA 法和Western blot 法检测多克隆抗体的效价和特异性。结果：ELISA 法检测所制 备的HPV16 E6、HPV16 E7 和HPV18 E7 多克隆抗体的效价可达1:2.56×105，HPV18 E6 多克隆抗体效价可达 1:1.28×105。Western blot 结果显示，所制备的抗体均能与相应的重组蛋白Flic-HPV16 E6E7 和Flic-HPV18 E6 E7 特异性结合。结论：采用人工合成肽作为免疫原成功制备了高效价、特异性好的兔抗HPV16 和HPV18 的E6 和E7 蛋白多克隆抗体，为后续进行HPV 感染检测、疫苗研发等实验研究创造了条件。

关键词: 人乳头瘤病毒, 多克隆抗体, E6 蛋白, E7 蛋白

Abstract:

Objective: To predict the B-cell linear epitopes of E6 and E7 proteins of human papillomavirus （HPV）16 and HPV18 and prepare rabbit polyclonal antibodies against E6 and E7 of HPV16 and HPV18. Methods：Bioinformatics software was used to analyze the amino acid sequences of E6 and E7 proteins of HPV16 and HPV18 to predict the B-cell linear epitopes. The peptides of HPV16 E6, HPV16 E7, HPV18 E6 and HPV18 E7 were designed and synthesized. The synthesized peptides were cross-linked with Keyhole Limpet Hemocyanin (KLH), which was used to immunize New Zealand white rabbits to prepare the polyclonal antibodies. Recombinant proteins HPV16 E6 E7 and HPV18 E6 E7 expressed in the prokaryotic system were used as antigens to detect the titer and specificity of rabbit polyclonal antibodies by indirect ELISA and Western blot. Results: The titers of HPV16 E6, HPV16 E7 and HPV18 E7 polyclonal antibodies were 1:2.56×105, and the titer of HPV18 E6 polyclonal antibody was 1:1.28×105. Western blot results showed that the antibodies could bind specifically to the recombinant proteins HPV16 E6 E7 and HPV18 E6 E7. Conclusion: The polyclonal antibodies against E6 and E7 of HPV16 and HPV18 were successfully prepared, which created the conditions for the subsequent experimental research on HPV infection or HPV vaccine development.

Key words: human papillomavirus, polyclonal antibodies, E6 protein, E7 protein

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