Development of an in process control filtration-assisted chemiluminometric immunoassay to quantify foot and mouth disease virus (FMDV) non-capsid proteins in vaccine-antigen batches.
In many countries, foot and mouth disease (FMD) is controlled by vaccination and surveillance against non-capsid proteins (NCP); therefore vaccines are required not to induce antibodies against NCP. Vaccine purity is evaluated by repeated inoculation of naïve cattle, an expensive and time consuming protocol that raises several animal welfare concerns. We have developed an in process control filtration-assisted chemiluminometric immunoassay (FAL-ELISA), to detect and quantify NCP in vaccine-antigen batches regardless of its volume and composition. Samples are filtered through PVDF-filter microplates pre-coated with a monoclonal antibody against NCP. Filtration removes all unbound components in the sample and captured NCP are detected by anti-NCP conjugate followed by incubation with the substrate, luminol/peroxide. Analytical detection limit was 2 ng for purified NCP and 4 ng for vaccine-antigen batches spiked with NCP, which makes this assay sensitive enough to be applied to purity control of FMD vaccines. Vaccine components did not interfere with the antibody and substrate reactions in the assay. FAL-ELISA is an alternative for the in vivo tests, observing the objective to Replace, Reduce and Refine the use of animals for quality control of immunobiologicals.
Capozzo
Alejandra Victoria
AV
Instituto de Ciencia y Tecnología Dr. César Milstein, Saladillo 2468, 1440 Ciudad de Buenos Aires, Argentina.
Martínez
Manuel Rosendo
MR
Schielen
Wilhelmus Joseph Gerardus
WJ
D055910:Antibodies, Immobilized / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005374:Filtration; D029544:Foot-and-Mouth Disease Virus / Q000737:chemistry*; D008163:Luminescent Measurements; D011786:Quality Control; D017361:Viral Nonstructural Proteins / Q000302:isolation & purification*; D014765:Viral Vaccines / Q000032:analysis*
C109754:3ABC protein, virus; D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines