| Ⅰ型单纯疱疹病毒单克隆抗体的制备及其应用; Preparation and application of monoclonal antibodies against Herpes simplex virus-1 | |
| 尹炳谦 ; 贾继宗 ; 赵风强 ; 韩金乐 ; 黄承浩 ; 叶祥忠 ; 张景海 | |
| 2015-06-20 | |
| 关键词 | HSV-1病毒 溶瘤 颗粒完整性 病毒滴度 HSV-1 Oncolytic Particle integrity Virus titer |
| 英文摘要 | 目的:制备并筛选HSV-1单抗,建立定量检测HSV-1病毒颗粒抗原的双抗体夹心ElISA方法,用于HSV-1病毒颗粒的质控。方法:以HSV-1免疫bAlb/C小鼠制备单克隆抗体,以筛选的中和单克隆抗体1f6为捕捉抗体,HrP标记的2b1为检测抗体,构建定量检测HSV-1病毒颗粒抗原的双抗体夹心ElISA方法,并对本方法的特异性、灵敏度、精密度、准确性和线性等性能进行验证。用本方法定量检测的病毒量与病毒滴度作回归分析。结果:构建的双抗体夹心定量检测HSV-1病毒颗粒抗原的ElISA方法,线性范围为0.125~2μg/Ml,相关系数为r2=0.995 5,定量限度为0.125μg/Ml,试剂的变异系数CV<10%,抗原回收率介于85.6%~107.1%之间。与HSV-1以外的其他样本无交叉反应。本方法检测与病毒感染滴度具有很好的相关性。结论:成功构建了定量检测HSV-1含量的ElISA方法,为HSV-1病毒颗粒抗原定量检测提供快速手段。; Objective: To prepare and screen monoclonal antibodies against Herpes simplex virus-1( HSV-1),and develop a double antibody sandwich quantitative enzyme-linked immunosorbent assay( Q-ELISA) for detection of HSV-1 particle.This method was used to control the quality of viral particle in the developing and manufacturing process of HSV-1.Methods: BALB / c mice was immunized with HSV-1 to prepare monoclonal antibodies.A double antibody sandwich Q-ELISA was developed to determine concentration of HSV-1 particle,which was based on the neutralizing monoclonal antibody 1F6 as capture antibody,and 2B1 as HRP-conjugated antibody.The performance of the reagent was evaluated,including specificity,sensitivity,precision,accuracy and linear.And the relation between the amount of virus detected by this method and the virus titer was analyzed by regression analysis method.Results: The QELISA for HSV-1 particle was developed.The quantitation scope was 0.125- 2 μg / ml,the coefficient correlation was 0.995 5,the limit of detection was 0.125 μg / ml,the recovery was between 85.6% and 107.1%,the variation coefficient was lower than 10%,and the reagent does not react with other samples except HSV-1 antigen.This method has a good correlation with virus titer.Conclusion: The QELISA for HSV-1 particle was successfully developed,which provide a new approach for rapid and quantitative detection of HSV-1 antigen. |
| 语种 | zh_CN |
| 内容类型 | 期刊论文 |
| 源URL | [http://dspace.xmu.edu.cn/handle/2288/119294]  |
| 专题 | 生命科学－已发表论文 |
| 推荐引用方式 GB/T 7714 | 尹炳谦,贾继宗,赵风强,等. Ⅰ型单纯疱疹病毒单克隆抗体的制备及其应用, Preparation and application of monoclonal antibodies against Herpes simplex virus-1[J],2015. |
| APA | 尹炳谦.,贾继宗.,赵风强.,韩金乐.,黄承浩.,...&张景海.(2015).Ⅰ型单纯疱疹病毒单克隆抗体的制备及其应用.. |
| MLA | 尹炳谦,et al."Ⅰ型单纯疱疹病毒单克隆抗体的制备及其应用".(2015). |
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