Preparation and application of monoclonal antibodies against Herpes simplex virus-1

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

Development of a quantitative ELISA detection method for Coxsackievirus A group 16 strain(CA16) antigen

目的:建立柯萨奇病毒A组16型(CA16)抗原的双抗体夹心ElISA定量检测方法,用于CA16灭活疫苗的研发和生产过程的抗原定量检测。方法:以CA16中和单抗T26H12为包被抗体、nA14b9为标酶抗体,构建定量检测CA16抗原的双抗体夹心ElISA方法,并对方法的特异性、灵敏度、精密度、准确性、线性和稳定性进行分析。结果:建立了双抗体夹心定量检测CA16抗原的ElISA方法。方法的线性相关系数r2=0.998,线性范围为8~128 ng/Ml,定量限度为8 ng/Ml;变异系数CV80%;与CA16以外的其他样本没有交叉反应。结论:构建的CA16抗原ElISA定量检测方法的各项性能符合定量检测需要,可用于CA16疫苗的研发和生产过程的抗原活性的定量检测。Objective:To develop an a quantitative enzyme linked immunosorbent assay(Q-ELISA) to determine the concentration of Coxsackievirus A Group 16 Strain(CA16) antigen.This method was used to determine CA16 antigen content at each stage of CA16 vaccine developing and manufacturing process.Methods:A double antibody sandwich Q-ELISA was developed to determine concentration of CA16 antigen,which was based on the high-affinity neutralizing monoclonal antibodies T26H12 as capture antibodies,and NA14B9 as HRP-labeled antibody.The performance of reagent were evaluated.Results:The Q-ELISA for CA16 antigen content was successfully developed.The reagent had good performance.The quantitation scope was 8-128 ng/ml,the coefficient correlation was 0.998,the limit of detection was 8 ng/ml,the recovery was between 87% and 113.8%.The stability was up to 80% after reagent was heated for 6 days at 37℃.The variation coefficient was lower than 15%,and thereagent was no reaction with other sample except CA16 antigen.Conclusion:The Q-ELISA for CA16 antigen was developed with good specificity,accuracy,precision and stability.The method can be used to determine CA16 antigen content during development and production of CA16 vaccine

Development of aquantitative ELISA detection method for Varicella Zoster Virus(VZV) antigen

目的:建立水痘-带状疱疹病毒(VzV)抗原的双抗体夹心ElISA定量检测方法,用于质控VzV灭活疫苗研发和生产中抗原含量。方法:以VzV中和单抗5f6C8为包被抗体、8H5d1为酶标抗体,构建定量检测VzV抗原的双抗体夹心ElISA方法,并对本方法的特异性、灵敏度、准确性、线性和稳定性等性能进行分析。结果:建立的双抗体夹心定量检测VzV抗原的ElISA方法,线性范围为0.4μg~13μg/Ml,相关系数为r2=0.994,定量限度为0.4μg/Ml;变异系数CV80%。与VzV以外的相关病毒样本没有交叉反应。结论:构建的VzV抗原ElISA定量检测方法的各项性能符合定量检测需要,可用于VzV灭活疫苗的研发和生产过程的抗原含量检测。Objective:To develop a quantitative enzyme linked immunosorbent assay(Q-ELISA) to determine the concentration of Varicella Zoster Virus(VZV) antigen.This method was used to determine VZV antigen content at each stage of VZV inactived vaccine developing and manufacturing process.Methods: A double antibody sandwich Q-ELISA was developed to determine concentration of VZV antigen,which was based on the the high-affinity neutralizing monoclonal antibodies 5F6C8 as capture antibodies,and 8H5D1 as HRP-labeled antibody.The performance of reagent were evaluated.Results: The Q-ELISA for VZV antigen content was successfully developed.The reagent had good performance.The quantitation scope was 0.4 μg~13 μg/ml,The coefficient correlation was 0.994,the limit of detection was 0.4 μg /ml,the recovery was between 87.5% and 111.6%.The stability was up to 80% after reagent was heated for 6 days at 37℃.The variation coefficient was lower than 15%,and the reagent was no reaction with other sample except VZV antigen.Conclusion: The Q-ELISA for VZV antigen was developed with good specificity,accuracy and stability.The method can be used to determine VZV antigen content during development and production of VZV inactived vaccine

Measurement of integrated luminosity of data collected at 3.773 GeV by BESIII from 2021 to 2024

We present a measurement of the integrated luminosity e+e- of collision data collected by the BESIII detector at the BEPCII collider at a center-of-mass energy of Ecm = 3.773 GeV. The integrated luminosities of the datasets taken from December 2021 to June 2022, from November 2022 to June 2023, and from October 2023 to February 2024 were determined to be 4.995±0.019 fb-1, 8.157±0.031 fb-1, and 4.191±0.016 fb-1, respectively, by analyzing large angle Bhabha scattering events. The uncertainties are dominated by systematic effects, and the statistical uncertainties are negligible. Our results provide essential input for future analyses and precision measurements

Determination of the number of ψ(3686) events taken at BESIII

The number of ψ(3686) events collected by the BESIII detector during the 2021 run period is determined to be (2259.3±11.1)×106 by counting inclusive ψ(3686) hadronic events. The uncertainty is systematic and the statistical uncertainty is negligible. Meanwhile, the numbers of ψ(3686) events collected during the 2009 and 2012 run periods are updated to be (107.7±0.6)×106 and (345.4±2.6)×106, respectively. Both numbers are consistent with the previous measurements within one standard deviation. The total number of ψ(3686) events in the three data samples is (2712.4±14.3)×10^