STANDARDIZATION OF POTENCY TEST METHODS FOR IPV-WHO STUDY

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 启动以 评估候选参比物质的适用性， 评估不同ELISA方法用于D测定的适用性， 并评估标准ELISA方法在不同条件下的变异性 实验室，使用相同的试剂和编码样本。 的材料 本研究中包括两种商业制备的候选参比品 材料;主要参考PU 78 -02目前使用的几个 实验室，来自不同制造商的疫苗显示， 免疫原性;从Sabin株和疫苗产生的IPV 体内效力差。 这项研究将分为两个阶段。 的 第一阶段已经完成，着眼于体外试验（ 第二阶段将着眼于体内测定）。 在CBER开发的ELISA 使用对D特异的单克隆抗体作为检测抗体 抗原是候选标准方法。 我们填充并冻干了我们的 分发给合作者的内部试剂。 一些测试是 在邮寄前进行验证冻干试剂。 我们 编写了详细的SOP，其中包括两种稀释方案 实践抗原和测试抗原。 合作者收到了一份 ELISA试剂盒，含有板、多克隆和单克隆抗体， 所有三种脊髓灰质炎病毒类型的单型抗原， 标准操作程序和我们用单型抗原的检测结果 标记为A-F的测试抗原在稍后的日期邮寄。 测试结果 目前正在评估。 我们计划 建议使用Balb/c小鼠作为候选标准体内效价方法 进行第二阶段的合作研究。 我们将参与 分析和报告每个阶段的研究。

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, assess the suitability of different ELISA methods for D determination, and to assess the variability of a standard ELISA method in different laboratories, using the same reagents and coded samples. The materials included in this study are two commercially prepared candidate reference materials; the primary reference PU78-02 currently used by several 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 has been completed, looked at in vitro assays (the second phase will look at 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. Test results have been compiled in a draft form and are being evaluated. We plan to propose using Balb/c mice as a candidate standard in vivo potency method for the second phase of this collaborative study. We will be involved in the analysis and reporting of the study at each phase.