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Method for the simultaneous assay of the different poliovirus types using surface plasmon resonance technology
Authors:Catherine Manin  Sophie NavilleMarine Gueugnon  Monique DupuyY Bravo de Alba  Olivier Adam
Institution:Sanofi Pasteur, Campus Mérieux, 1541 Avenue Marcel Mérieux, 69280 Marcy l’Etoile, France
Abstract:The inactivated polio vaccine (IPV) contains viral samples that belong to serotypes 1, 2 and 3. We report here a surface plasmon resonance (SPR)-based technique that permits the simultaneous assay of the individual viral types in the IPV as well as in different bulk intermediates from the industrial vaccine production process. Monoclonal antibodies specific to each of the 3 viral types along with a negative control antibody are captured via an anti-IgG antibody on the surface of the 4 flow cells of the SPR instrument. The viral samples are then injected over these flow cells and the increase in resonance units as a result of virus binding is measured. The method was calibrated by an analysis of the European Working Standard (EWS) for poliovirus vaccines. We show that the antibodies used recognize viruses with functional affinities in the picomolar range permitting an effective capture of the antigen. In addition we demonstrate that the antibodies are highly specific to a given virus type and that the heat induced destruction of the D-antigen abolishes antibody recognition entirely. The technique was found to be reproducible and robust and its response was linear to the antigen concentration. Due to the rapidity of analysis this technique permits an almost real-time follow-up of the industrial production process and may present an alternative to the established ELISA assay for the analysis of the intermediates and the final product.
Keywords:EDC  1-ethyl-3-(3-dimethylpropyl)-carboiimide  DU  D-antigen unit  ELISA  enzyme-linked immunosorbent assay  EWS  European Working Standard  HBS-EP  hepes-buffered saline containing EDTA and the P20 detergent  Ig  immunoglobulin  IPV  inactivated polio vaccine  Mab  monoclonal antibody  MCK  multi-cycle kinetic  PV  poliovirus  NHS  N-hydroxysuccinimide  OPV  oral polio vaccine  SCK  single-cycle kinetic  SPR  surface plasmon resonance  PV  poliovirus  RU  resonance unit
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