应用噬菌体展示制备人乳头瘤病毒6型L1蛋白中和性单克隆抗体
Preparation of neutralizing monoclonal antibodies against human papillomavirus type 6 L1 protein using phage display
秦薇子 1陈桂榴 1张黎2
作者信息
- 1. 中国科学技术大学附属第一医院(安徽省立医院),合肥 230001
- 2. 江苏省疾病预防控制中心,南京 210009
- 折叠
摘要
目的 制备人乳头瘤病毒6型L1(HPV6L1)蛋白特异性单克隆抗体,探索抗体的结合特点、交叉活性以及中和效果.方法 从免疫HPV6 L1蛋白小鼠脾细胞中提取抗体基因,构建ScFv噬菌体抗体文库.用HPV6 L1蛋白对文库进行富集筛选,将获得的抗体基因表达成鼠IgG后,验证抗体与L1蛋白的结合性质以及对假病毒的中和效果.结果 成功构建了针对HPV6 L1的ScFv抗体文库,库容量为6.54×107.抗体文库经HPV6 L1筛选后获得了两株抗体,命名为Q9和Q11.经ELISA检测,Q9-IgG与HPV6 L1反应的滴度约为106,与HPV18和45 L1的滴度为102.Q11-IgG与HPV6 L1结合的滴度约为4×104,而与其他型别L1均不结合.经过假病毒中和试验测定,Q9-IgG没有中和活性,而Q11-IgG针对HVP6假病毒的中和滴度约为104.5.结论 Q9和Q11均为HPV6 L1特异性单克隆抗体,但二者交叉反应性、抗原识别位点及中和活性均具有明显差异.该抗体可为HPV6病毒的基础研究、免疫学诊断以及治疗制剂研发提供工具抗体.
Abstract
Objective To prepare monoclonal antibodies specific to human papillomavirus type 6 L1(HPV6 L1)protein,investigate the binding characteristics,cross-reactivity,and neutralization efficacy of these antibodies.Methods Antibody genes were extracted from the spleen cells of mice immunized with HPV6 L1 protein and used to construct an ScFv phage antibody library.The library was enriched and screened using HPV6 L1 protein.Following expression of the antibody genes into murine IgG,the binding properties of the antibodies to the L1 protein and their neutralization effects on pseudoviruses were verified.Results An ScFv antibody library targeting HPV6 L1 was successfully constructed with a capacity of 6.54x 107.After screening with HPV6 L1,two antibody strains were obtained,named Q9 and Q11.ELISA testing revealed that Q9-IgG has a high antibody titer(about 106)against HPV6 L1,while cross-reactivity titers to HPV18 and 45 L1 were 102.Q11-IgG has an antibody titer about 4×104 against HPV6 L1,but not combined with the L1 proteins of other HPV types.Pseudovirus neutralization assays indicated that Q9-IgG lacked neutralizing activity,while Q11-IgG showed a neutralizing antibody titer of 104.5 against HPV6 pseudovirus.Conclusions Both Q9 and Q11 are monoclonal antibodies specific to HPV6 L1.However,they displayed notable differences in cross-reactivity,antigen recognition sites,and neutralization efficacy.These antibodies provide essential tools for the foundational research of HPV6,immunological diagnosis,and the development of therapeutic formulations.
关键词
人乳头瘤病毒6型/L1蛋白/噬菌体展示/单克隆抗体Key words
Human papillomavirus type 6/L1 protein/Phage display/Monoclonal antibody引用本文复制引用
出版年
2024
NETL
NSTL
万方数据