STANDARDIZATION OF POTENCY TEST METHODS FOR IPV-WHO STUDY

当前脊髓灰质炎灭活病毒疫苗(IPV)效力测试需要 实验室之间的标准化。一项合作研究已经完成 启动以评估候选参考材料的适宜性， 评价不同的酶联免疫吸附试验方法测定D的适用性， 并评估标准的酶联免疫吸附试验方法在不同的 实验室，使用相同的试剂和编码样本。这些材料 这项研究包括两个商业准备的候选参考文献 材料；目前由几个人使用的主要参考PU78-02 实验室，来自不同制造商的疫苗显示 在人类中具有免疫原性；由沙宾病毒株和疫苗产生的IPV 体内效力差。这项研究将分为两个阶段。这个 第一阶段已经完成，观察了体外分析( 第二阶段将着眼于体内分析)。CBER研制的一种酶联免疫吸附试验 用D特异性单抗作为检测抗体的研究 抗原是候选标准方法。我们装满并冷冻干燥我们的 分发给合作者的内部试剂。有几项测试是 在邮寄前对冻干试剂进行验证。我们 准备了一份详细的SOP，其中包括两种情况下的稀释方案 练习抗原和测试抗原。合作者收到了一个酶联免疫吸附试验 单型多克隆和单克隆抗体试剂盒 用于实践检测的所有三种脊髓灰质炎病毒类型的抗原 加上标准操作规程和CBER对实践抗原的结果。 编码的测试抗原随后被邮寄。尽管做出了这些努力， 抗体效价的检测结果令人无法接受。 不同实验室之间的可变性。此外，类似的 在比较效力测定时注意到了可变性 内部酶联免疫吸附测定。这项合作研究的结果 (Wood，D.J.，Heath，A.B.and Sawyer，L.A.，Biologics，1995；23：83-94 已经表明有必要进一步提高效力 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.