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 开始评估候选参考材料的适用性， 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ELISA 含有平板、多克隆和单克隆抗体、单型 所有三种脊髓灰质炎病毒类型的抗原， 除了SOP和实践抗原的CBER结果。 编码 随后邮寄测试抗原。 尽管作出了这些努力， 单克隆ELISA试验的效价结果显示不可接受 不同实验室之间的差异。 此外，类似 在比较基于以下的效价测定结果时注意到变异性： 内部ELISA测定。 这项合作研究的结果 (Wood，D.J.，Heath，A.B.洛杉矶的索耶生物制品，1995;23：83-94 已经表明需要进一步提高效力 IPV的确定。

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. Collaborators received an ELISA kit containing plates, polyclonal and monoclonal antibodies, monotypic antigens of all three poliovirus types for practice determinations in addition to the SOPs and the CBER results for the practice antigens. Coded test antigens were subsequently mailed. Despite these efforts, the potency results for the monoclonal ELISA assay exhibited unacceptable variability between different laboratories. Furthermore, similar variability was noted in comparing potency determinations based on in-house ELISA determinations. The results of this collaborative study (Wood, D.J., Heath, A.B. and Sawyer, L.A., Biologicals, 1995;23:83-94 have indicated the need for further improvements in potency determinations of IPV.