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Demonstrates one-step immunoaffinity purification of Bordetella pertussis filamentous haemagglutinin using monoclonal antibodies, highlighting an optimized detection and isolation strategy.
Two monoclonal antibodies against filamentous haemagglutinin (F-HA) from Bordetella have been produced. In immunoblotting of a SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of high ionic strength extracts of Bordetella pertussis, the two monoclonal antibodies both stained several high molecular weight bands. The two monoclonal antibodies were used for immuno-affinity column one-step purification of F-HA from high-ionic-strength extracts of Bordetella pertussis. In gradient SDS-PAGE, the eluted F-HA is present as a major double band with a molecular weight of approximately 240,000 daltons and several minor bands with molecular weights down to 90,000 daltons. The monoclonal antibodies were used in a monoclonal antibody catching enzyme linked immunosorbent assay (ELISA) for sensitive and specific detection of F-HA. This ELISA system proved valuable in optimizing the elution conditions for the monoclonal affinity columns.
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Selmer et al. (2009) studied this question.
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