Abstract：
: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.
journal_name
Vaccinejournal_title
Vaccineauthors
Capozzo AV,Martínez MR,Schielen WJdoi
10.1016/j.vaccine.2010.05.049subject
Has Abstractpub_date
2010-09-14 00:00:00pages
6647-52issue
40eissn
0264-410Xissn
1873-2518pii
S0264-410X(10)00747-4journal_volume
28pub_type
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