Comparison of Three Independently Produced Alternaria r Alt a1

RATIONALE: Alternaria Alt a1 allergen has been identified as a major Alternaria allergen. Quantitative assays has been developed to detect Alt a1 in allergenic extracts for standardization, and environmental samples for exposure assessment and monitoring the efficacy of avoidance measures. These assays are critical in the understanding of Alternaria sensitivity.METHODS: In this study we sought to compare the monoclonal antibody and Human IgE binding profile to three independently produced recombinant Alt a1. rAlt a1 was secured from three different sources, Bial-Aristegui, Spain, Indoor Biotechnologies, VA, USA, and our own laboratory. The material was quantitated and an equal amount of rAlt a1 was Electrophoresed in a 15 % SDS-PAGE, then transferred onto PVDFmembrane.RESULTS: A murine monoclonal antibody was used to probe the membrane for binding to rAlt a1. We also probed with IgE from pooled sera from 15 Alternaria sensitive individuals. Both blots showed antibody binding to Alt a1 but at different intensities.CONCLUSION: We concluded that the manner in which r Alt a1 is produced and purified may affect its biological activity. RATIONALE: Alternaria Alt a1 allergen has been identified as a major Alternaria allergen. Quantitative assays has been developed to detect Alt a1 in allergenic extracts for standardization, and environmental samples for exposure assessment and monitoring the efficacy of avoidance measures. These assays are critical in the understanding of Alternaria sensitivity. METHODS: In this study we sought to compare the monoclonal antibody and Human IgE binding profile to three independently produced recombinant Alt a1. rAlt a1 was secured from three different sources, Bial-Aristegui, Spain, Indoor Biotechnologies, VA, USA, and our own laboratory. The material was quantitated and an equal amount of rAlt a1 was Electrophoresed in a 15 % SDS-PAGE, then transferred onto PVDFmembrane. RESULTS: A murine monoclonal antibody was used to probe the membrane for binding to rAlt a1. We also probed with IgE from pooled sera from 15 Alternaria sensitive individuals. Both blots showed antibody binding to Alt a1 but at different intensities. CONCLUSION: We concluded that the manner in which r Alt a1 is produced and purified may affect its biological activity.