夏百成,刘洪洋,郝亚茹,张 婷,马铭饶,许雪梅.人乳头瘤病毒(HPV)31型中和单克隆抗体的制备及性质鉴定[J].,2020,(6):1001-1005 |
人乳头瘤病毒(HPV)31型中和单克隆抗体的制备及性质鉴定 |
Preparation and Characterization of Neutralizing Monoclonal Antibodies Against Human Papillomavirus Type 31 |
投稿时间:2019-09-27 修订日期:2019-10-23 |
DOI:10.13241/j.cnki.pmb.2020.06.001 |
中文关键词: 人乳头瘤病毒31型 L1病毒样颗粒 单克隆抗体 中和活性 |
英文关键词: Human papillomavirus Virus-like particle Monoclonal antibody Neutralizing activity |
基金项目:中国医学科学院医学与健康科技创新工程项目基金 (2016-I2M-3-026);国家国际科技合作项目 (2013DFA32430) |
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中文摘要: |
摘要 目的:构建获得HPV31构象依赖的中和单抗。方法:采用昆虫细胞表达的HPV31 L1VLP(virus-like particle, VLP)免疫BALB/c小鼠,取免疫鼠脾细胞与SP2/0细胞融合,收集杂交瘤细胞培养上清,首先采用HPV31 VLP-ELISA及HPV31假病毒中和实验筛选分泌HPV31中和单抗的杂交瘤细胞株,然后纯化单克隆抗体,分别采用假病毒中和实验及ELISA实验对纯化后单克隆抗体进行鉴定,包括抗体的亚型及其结合表位的构象特征、针对HPV31的中和IC50及针对HPV16、HPV18、HPV33、HPV45、HPV52、HPV58、HPV6、HPV11的交叉中和活性。结果:筛选获得9株HPV31构象依赖中和单抗,其中3株HPV31特异性中和单抗中有2株的 IC50<10 ng/mL,分别是XM31-13(0.36,IgG1)及XM31-23(7.10,IgG1),6株交叉中和单抗中亦有2株的 IC50<10 ng/mL,分别是XM31-19(7.14, IgG1)、XM31-20(6.91,IgG1)。结论:获得的9株HPV31构象依赖的中和单抗,特别是其中4株单抗IC50值 < 10 ng/mL的4株单抗,可用于含HPV31L1VLP多价疫苗的质控疫苗的研究。 |
英文摘要: |
ABSTRACT Objective: Preparation of some conformation-dependent neutralizing monoclonal antibodies (mAbs) against HPV31. Methods: Female BALB/c mice were immunized with HPV31 L1 virus-like particles (VLPs) expressed by insect cells, and spleenocytes of the zimmunized mice were fused with SP2/0 cells to prepare hybridoma cells, the culture supernatants were-collected for identification of neutralizing mAb secreting cells by HPV31 VLP-ELISA and HPV31 pseudovirus neutralization experiments. Monoclonal antibodies were purified and identified by ELISA and HPV31, HPV16, HPV18, HPV52, HPV58, HPV33, HPV45, HPV6 or HPV11 pseudovirus neutralization experiments. Results: Nine HPV31 conformation-dependent neutralizing mAbs were screened, and three of which were HPV31 specific mAbs and the other six were cross-neutralizing mAbs.The mAbs whith IC50<10 ng/mL were XM31-13 (0.36), XM31-19 (7.14), XM31-20 (6.9), XM31-23 (7.10). Conclusion: Nine conformation-dependent neutralizing mAbs were obtained, and four of them have low IC50 value, which is useful for development of HPV31-VLP containing multivalemt vaccines. |
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