Sawyer L A, McInnis J, Albrecht P
Laboratory of Pediatric Diseases, Center for Biologics Evaluation and Research, Rockville, Maryland 20852-1448.
Biologicals. 1993 Jun;21(2):169-77. doi: 10.1006/biol.1993.1070.
The enzyme-linked immunosorbent assay (ELISA) is currently the in vitro method for the measurement of the D antigen content of inactivated poliovirus vaccines (IPV) of greatest interest. The sensitivity and specificity of the test is dependent on the antibodies selected for use. We evaluated monoclonal and polyclonal antibodies for specificity to D antigen and wild or attenuated (Sabin) strains used in vaccine production. When used as detection antibodies the types 1 and 2 monoclonal antibodies raised against wild-type poliovirus strains were D antigen-specific and cross-reactive with the corresponding Sabin strains. The type 3 monoclonal antibody was weakly cross-reactive with Sabin type 3 vaccine. In contrast, polyclonal antibodies were less D antigen-specific, but reacted equally well with wild-type and Sabin strain vaccines. The ELISA using monoclonal antibodies was shown to be highly reproducible. Reactivity with these monoclonal antibodies implies that a D-specific neutralizing epitope of each respective poliovirus type has been preserved in the inactivation process. Evaluation with additional neutralizing D antigen-specific monoclonal antibodies may be necessary to determine whether reactivity with one epitope of each type in an in vitro test is sufficient to predict potency of the vaccine in humans.
酶联免疫吸附测定(ELISA)是目前用于测量灭活脊髓灰质炎疫苗(IPV)中D抗原含量的最受关注的体外方法。该检测方法的灵敏度和特异性取决于所选用的抗体。我们评估了针对D抗原以及疫苗生产中使用的野生型或减毒(萨宾)毒株的单克隆抗体和多克隆抗体的特异性。当用作检测抗体时,针对野生型脊髓灰质炎病毒株产生的1型和2型单克隆抗体具有D抗原特异性,并且与相应的萨宾毒株有交叉反应。3型单克隆抗体与萨宾3型疫苗的交叉反应较弱。相比之下,多克隆抗体的D抗原特异性较低,但与野生型和萨宾毒株疫苗的反应同样良好。使用单克隆抗体的ELISA显示出高度的可重复性。与这些单克隆抗体的反应性表明,每种脊髓灰质炎病毒型别的D特异性中和表位在灭活过程中得以保留。可能需要用更多的D抗原特异性中和单克隆抗体进行评估,以确定在体外试验中与每种类型的一个表位的反应性是否足以预测疫苗在人体中的效力。