High-Throughput Method for Determining the Presence of Papillomavirus-Neutralizing Antibodies in a Sample
The invention is based on papillomavirus pseudovirions consisting of L1 and L2 proteins encapsulating Gaussia luciferase as a reporter system. Using HeLaT K4 cells as target cells the screen can be performed in a high-throughput format without the need of time consuming manual pipetting steps. The method
Cervical cancer is the second leading course of cancer-related death in women, causing approximately 250,000 deaths each year. Given the fact that infection with HPV viruses is evident in almost all cases and that vaccination against high-risk HPV viruses has been proven to be a valuable tool for cancer prevention, methods are needed for determining the HPV immune status. The present invention offers a screen for HPV antibodies based on pseudovirion neutralization and which can be used in a high-throughput format.
Currently, two methods for determining titers of neutralizing antibodies against HPV are available in the art: antibody competition assays and assays based on pseudovirion neutralization. While the former suffers from drawbacks related to insufficient correlation between vaccination infectivity and measured antibody levels, the latter involves timeconsuming manual pipetting steps. The current invention is therefore based on papillomavirus pseudovirions consisting of L1 and L2 proteins encapsulating Gaussia luciferase as a reporter system. After incubation of the pseudovirions with e.g. patient serum, HeLaT K4 cells are added to the particle-serum mix. Cells are cultivated for 48h and the amount of secreted luciferase is subsequently determined by luminescence. At the moment antibodies against thirteen high-risk HPV serotypes can be detected.
- Test for functional antibodies against HPV L1 and L2 proteins based on pseudovirion neutralization assa
- Exact quantitative and qualitative determination of antibody titer in sample fluid in high-throughput format
- HT test is suitable for different types of body fluids and many HPV subtypes
- Technology opportunity is available as service or non-exclusive license
The system can be used for quantitative and qualitative determination of HPV αL1 and αL2 antibody titers for most subtypes of HPV viruses in a highthroughput format.
Publikationen & Verweise
- Sehr et al., PLoS One. 2013 Oct 4;8(10). See at: https://www.ncbi.nlm.nih.gov/pubmed/?term=High-Throughput+Pseudovirion-Based+Neutralization+Assay+for+Analysis+of+Natural+and+Vaccine-Induced+Antibodies+against+Human+Papillomaviruses
- Toft et al., J Infect Dis. 2013 Dec 26. See at: https://www.ncbi.nlm.nih.gov/pubmed/?term=Comparison+of+the+Immunogenicity+and+Reactogenicity+of+Cervarix+and+Gardasil+Human+Papillomavirus+Vaccines+in+HIV-Infected+Adults%3A+A+Randomized%2C+Double-Blind+Clinical+Trial
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StichworteDiagnostics, Research Tools, Vaccines