STANDARDIZATION OF POTENCY TEST METHODS FOR IPV-WHO STUDY

目前灭活脊髓灰质炎病毒疫苗（IPV）效力的测试需要 实验室之间的标准化。 一项合作研究已 开始评估候选参考材料的适用性， 评估不同 ELISA 方法对 D 测定的适用性， 并评估标准 ELISA 方法在不同环境中的变异性 实验室，使用相同的试剂和编码样本。 材料 本研究包括两个商业准备的候选参考 材料；主要参考 PU78-02 目前被几个人使用 实验室显示，来自不同制造商的疫苗 人类具有免疫原性；由 Sabin 毒株和疫苗生产的 IPV 体内效力较差。 该研究将分为两个阶段。 的 第一阶段已经完成，研究了体外测定（ 第二阶段将着眼于体内测定）。 CBER 开发的 ELISA 使用D特异性单克隆抗体作为检测抗体 抗原是候选标准方法。 我们填充并冻干我们的 内部试剂分发给合作者。 进行了多项测试 在邮寄前进行验证冻干试剂。 我们 准备了详细的 SOP，其中包括两者的稀释方案 练习抗原和测试抗原。 合作者接受了 ELISA 包含板、多克隆和单克隆抗体的试剂盒，单型 所有三种脊髓灰质炎病毒类型的抗原，用于实践测定 除了实践抗原的 SOP 和 CBER 结果之外。 随后邮寄了编码的测试抗原。 尽管做出了这些努力， 单克隆 ELISA 测定的效力结果显示不可接受 不同实验室之间的差异。 此外，类似的 在比较基于以下因素的效力测定时注意到了变异性 内部 ELISA 测定。 这项合作研究的结果 （Wood, D.J.、Heath, A.B. 和 Sawyer, L.A.，生物制品，1995 年；23:83-94 表明需要进一步提高效力 IPV 的测定。

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.