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Immunochemical and physicochemical characterization of commercial Altemaria extracts: a model for standardization of mold allergen extracts
Authors:MK Agarwal  RT Jones  JW Yunginger
Institution:1. From the Departments of Pediatrics and Internal Medicine (Allergy), Mayo Medical School, Rochester, Minn., USA;2. From the Allergic Diseases Research Laboratory, Mayo Clinic and Foundation, Rochester, Minn., USA
Abstract:To develop a model for mold allergen extract standardization, we studied eight commercial Alternaria extracts from various suppliers by a variety of immunochemical and physicochemical techniques, including measurement of Alt-I, a purified allergenic fraction of Alternaria. Wide variations were noted in the allergenic and antigenic potencies of these extracts. Estimates of Alt-I content measured by Alt-I RAST inhibition and by radioimmunoassay correlated significantly (p < 0.05), but Alt-I activity by either method could not be correlated with allergenic potency as measured by RAST inhibition using solid-phase Alternaria. Each test extract produced unique and differing patterns of Coomassie blue-stained bands in isoelectrofocusing gels and in crossed immunoelectrophoresis gels using rabbit antibodies to Alternaria. The optimal method for mold allergen standardization involves a combination of RAST inhibition, isoelectrofocusing, and crossed immunoelectrophoresis techniques, and, if possible, quantitation of individual allergens.
Keywords:CIE  Crossed immunoelectrophoresis  CRIE  Crossed radioimmunoelectrophoresis  DARIA  Double-antibody radioimmunoassay  PNU  Protein nitrogen unit  TLIEF  Thin-layer isoelectrofocusing  TNU  Total nitrogen unit
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