Process Control Filtration-Assisted Chemiluminometric Immunoassay to Quantify Foot and Mouth Disease virus (FMDV) Non Capsid Proteins in Vaccine Antigen Batches

CAPOZZO, A.V.1*, MARTÍNEZ, M1.; AND SCHIELEN, WJG

SENASA Martinez

Simposio; Practical Alternatives to Reduce Animal Testing in Quality Control of Veterinary Biologicals in the Americas; 2010

OIE - Fundación PROSAIA

Commercial FMDV vaccine antigenic preparations have to be partially purified to remove non-capside proteins (NCP, particularly 3ABC), involved in the replication of the virus, in order to allow serological testing to differentiate between infected and vaccinated animals (DIVA). Determination of 3ABC in vaccine batches is actually done in vivo, by serology testing of naïve cattle. Here we describe the development of an in process control, filtration-assisted luminometric immunoassay, to detect and quantify 3ABC in viral-vaccine batches. Aliquots of known sample size are filtered through PVDF-filter microplates pre-coated with a monoclonal antibody (Mab) against 3ABC. Filtration cleans up the sample and large volumes can be used. 3ABC captured by the specific Mab can be visualized by anti-3ABC HPRT conjugate followed by luminol/peroxide. Analytical detection limit is about 2 ng of 3ABC per vaccine dose, what makes it sensitive enough to be applied to purity control of FMD vaccines. No reaction has been detected with confirmed negative samples. This assay allows superior low-end sensitivity and faster protocol than conventional colorimetric methods, plus the versatility of testing any sample, regarding its particular composition.