Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus...

Area of Science:

• Immunology
• Vaccinology
• Analytical Chemistry

Background:

• Multiplex immunoassays are crucial for assessing antibody responses in 9-valent human papillomavirus (9vHPV) vaccine clinical trials.
• The HPV6/11/16/18/31/33/45/52/58 competitive Luminex immunoassay (HPV-9 cLIA) and total immunoglobulin G Luminex immunoassay are routinely used for immunogenicity measurements.
• Assay modernization is vital for vaccine life cycle management.

Purpose of the Study:

• To redevelop and validate two multiplex immunoassays for measuring antibody responses to the 9vHPV vaccine.
• To ensure continuity and comparability of immunogenicity data by bridging redeveloped assays to previous versions.
• To establish accurate and precise assays suitable for high-throughput testing in current and future HPV vaccine studies.

Main Methods:

• Optimization of key assay parameters including coating concentration, Luminex microspheres, diluents, and antibody conjugates.
• Validation of redeveloped assays for performance characteristics such as precision, linearity, accuracy, and quantitation limits.
• Bridging studies using patient sera from a 9vHPV vaccine clinical trial to link new assay versions with historical data.

Main Results:

• Both redeveloped immunoassays demonstrated accuracy, specificity, and precision within their quantifiable ranges.
• Strong linear associations were observed between the new and previous versions of both assays.
• Assay serostatus cutoffs were established and refined to ensure alignment of seropositivity rates between assay versions.

Conclusions:

• The reoptimized 9vHPV immunoassays are validated and suitable for use in ongoing and future clinical trials.
• Bridging to previous assay versions allows for continuous monitoring of immune responses across different study phases and populations.
• These modernized assays support high-throughput assessment of HPV antibodies, facilitating vaccine efficacy and safety monitoring.