STANDARDIZATION OF POTENCY TEST METHODS FOR IPV-WHO STUDY

IPV-WHO 研究效力测试方法的标准化

• 批准号: 3770312
• 负责人: L A SAWYER
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3770312
• 关键词: World Health Organization; drug quality /standard; enzyme linked immunosorbent assay; laboratory mouse; lyophilization; monoclonal antibody; nonhuman therapy evaluation; poliomyelitis vaccine; poliovirus; reagent /indicator; virus antigen

The current tests for inactivated poliovirus vaccine (IPV) potency need standardization between laboratories. A collaborative study has been initiated to assess the suitability of candidate reference materials. Another aspect of the study is to assess the suitability of a candidate standard method for D antigen assay by ELISA, and to assess the proficiency of methods for potency estimation of IPV used by individual laboratories. The materials included in this study are two commercially prepared candidat reference materials; the primary reference PU78-02 currently used by severa laboratories, vaccines from different manufacturers shown to be immunogenic in humans; IPV produced from Sabin strains and vaccines of poor in vivo potency. The study will be divided into two phases. The first phase which is in progress will involve in vitro assays and the second phase in vivo assays. An ELISA developed at CBER using as detection antibodies monoclonal antibodies specific for D antigen is the candidate standard method. We filled and lyophilized our in-house reagents for distribution to collaborators. Several tests were performed to validate the lyophilized reagents prior to mailing. We prepared a detailed SOP which included the dilution schemes for both practice antigens and test antigens. The collaborators have received an ELISA kit containing plates, polyclonal and monoclonal antibodies and monotypic antigen of all three poliovirus types for practice together with the SOPs and the results of our tests using the monotypic antigens. The test antigens labeled A-F were mailed at a later date. Testing is in progress. We are currently evaluating the Balb/c mous as a model of in vivo potency as a separate study. We plan to propose usin Balb/c mice as a candidate standard in vivo potency method for the second phase of the collaborative study. We will be involved in the analysis and reporting of the study at each phase.

目前，灭活脊髓灰质炎病毒疫苗（IPV）效力的检测需要 实验室之间的标准化。 一项合作研究已经 开始评估候选参考物质的适用性。 这项研究的另一个方面是评估候选人的合适性 ELISA法检测D抗原的标准方法，并评估熟练程度 各实验室使用的IPV效价估计方法。 本研究中包括的材料是两种商业制备的候选材料， 标准物质;目前由几家公司使用的主要标准物质PU 78 -02 实验室，来自不同制造商的疫苗显示具有免疫原性 在人类中;由Sabin株产生的IPV和体内不良 力量 这项研究将分为两个阶段。 第一阶段， 正在进行的第二阶段将包括体外试验和体内试验 分析。 在CBER开发的ELISA使用单克隆抗体作为检测抗体 D抗原特异性抗体是候选标准方法。 我们 灌装和冻干我们的内部试剂，以分发给 合作者 进行了几项测试以验证冻干的 在邮寄之前， 我们准备了详细的SOP，其中包括 用于实践抗原和测试抗原的稀释方案。 的 合作者已经收到了含有板、多克隆和 所有三种脊髓灰质炎病毒的单克隆抗体和单型抗原 与SOP和我们使用的测试结果一起进行实践 单型抗原 标记为A-F的测试抗原稍后邮寄给 约会 测试正在进行中。 我们目前正在评估Balb/c小鼠 作为单独研究的体内效力模型。 我们计划建议使用 Balb/c小鼠作为第二种体内效价方法的候选标准 合作研究阶段。 我们将参与分析， 每个阶段的研究报告。