| 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].现代生物医学进展英文版,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 |
| Received:September 27, 2019 Revised:October 23, 2019 |
| 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) |
| Author Name | Affiliation | E-mail | | XIA Bai-cheng | Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100005, China | xiabaicheng1@126.com | | LIU Hong-yang | Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100005, China | | | HAO Ya-ru | Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100005, China | | | ZHANG Ting | Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100005, China | | | MA Ming-rao | Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100005, China | | | XU Xue-mei | Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100005, China | |
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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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