Generation of Monoclonal Antibodies and Development of Sandwich ELISA for 9-valent HPV Vaccines

Cervical cancer is one of the most common cancers affecting women globally, and nearly all cases of cervical cancer were caused by human papillomavirus (HPV) infection. HPV vaccination has proven to be effective strategy in preventing HPV infection. In this study, for the nine-valent HPV vaccines (HPV 6, 11, 16, 18, 31, 33, 45, 52, and 58), we have generated subtype-specific mouse monoclonal antibodies for all nine HPV subtypes by immunizing the mice with using 9 different virus-like particle (VLP) respectively. Neutralizing monoclonal antibodies specific against each HPV subtypes were identified and used to develop the Sandwich ELISA kits. They can specifically measure the concentration of each subtype VLP in the mixture of 9-valent HPV vaccine for drug substance (DS) or drug product (DP) samples. Most importantly, the ELISA kits can detect the temperature-induced loss of conformational neutralizing antigen epitope(s) in the vaccines, thereby serving as a tool could be used to monitor the stability of the 9-valent HPV vaccines in vitro. In addition, these 9 HPV subtype VLP Sandwich ELISA kits are compatible with HPV VLP vaccines produced by six different vaccines manufacturers, which use three different expression systems with or without the disassembling/reassembling process. Highlights Subtype-specific HPV VLP Sandwich ELISA kits for the nine-valent HPV vaccines (HPV 6, 11, 16, 18, 31, 33, 45, 52, and 58) were developed using neutralizing monoclonal antibodies These kits can specifically measure the concentration of each subtype VLP in the mixture of 9-valent HPV vaccine for drug substance (DS) or drug product (DP) samples. They can detect the temperature-induced loss of conformational neutralizing antigen epitope(s) in the vaccines, thereby serving as a tool to monitor the stability of the 9-valent HPV vaccines in vitro. These ELISA kits are compatible with HPV VLP vaccines produced by six different vaccines manufacturers, which use three different expression systems with or without the disassembling/reassembling process.