Generation of Polyclonal Antibodies Against Sabin Poliovirus D- and H-Antigens and Their Application in ELISA

Background/Objectives: The World Health Organization (WHO) recommends the use of attenuated Sabin strains for the production of inactivated poliovirus vaccine (IPV), offering improved biosafety while retaining immunogenicity. To better characterize the antigenic composition of Sabin strain-based IPV (sIPV), including both the protective D-antigen and the non-protective H-antigen forms, we developed a method for purifying D- and H-antigens forms. Methods: D- and H-antigens of poliovirus Sabin strains types 1, 2, and 3 were purified using gradient ultracentrifugation and used to generate antigen-specific polyclonal antibodies. Results: The generated polyclonal antibodies demonstrated high specificity with neutralizing titers of antibodies against Sabin type 1 poliovirus—1:2048, against Sabin type 2 poliovirus—more than 1:2048, against Sabin type 3 poloivirus —1:2048. Conclusions: This antigen-specific antibody approach provides a valuable tool for routine quality control in sIPV manufacturing, enabling accurate quantification of immunogenic components and detection of potentially immunogenic degradation products during vaccine storage and distribution. Antibodies to the D-antigen allow assessment of immunogenic, neutralizing epitopes, while antibodies to the H-antigen provide a tool for detecting non-neutralizing components. This antigen-specific antibody approach offers a valuable tool for studying the antigenic structure of sIPV and for improving the accuracy of ELISA-based antigen quantification.