M L Patterson1, J E Slater. 1. Laboratory of Immunobiochemistry, Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Bethesda 20852, USA. pattersonm@cber.fda.gov
Abstract
BACKGROUND: In this study we examine the variability among unstandardized cockroach allergen extracts. METHODS: We obtained 24 aqueous and glycerinated cockroach allergen extracts from nine manufacturers. We used previously characterized cockroach extracts, E2-Cg and E2-Ca, as references. The modified ninhydrin assay was used to determine protein concentration of each extract. Relative potencies of extracts were determined by competition ELISA, using a human allergic serum pool. Bla g 1 and Bla g 2 levels of glycerinated German cockroach extracts were determined by ELISA using monoclonal antibodies. Extracts were also analysed by SDS-PAGE. RESULTS: Commercial cockroach allergen extracts had highly variable protein contents that were lower than the protein contents of the references. Electrophoretic data confirmed the presence of a variable number and intensity of protein bands in extracts among manufacturers. The relative potencies of the commercial extracts were between 10 and 782 BAU/mL for German cockroach and 10-250 BAU/mL for American cockroach. The mean Bla g 1 content of the commercial extracts was significantly lower than that of the reference (P = 0.001). The mean Bla g 2 content of the commercial extracts was higher than that of the E2-Cg reference but the Bla g 2 levels were more variable compared to Bla g 1. In glycerinated German cockroach extracts, protein concentrations, relative potencies and specific allergen levels were significantly correlated (P < 0.001). CONCLUSION: Our tests indicate that commercially available cockroach allergen extracts are variable in protein content, electrophoretic banding patterns, relative potency and Bla g 2 levels. In glycerinated German cockroach extracts, protein concentrations, relative potencies and specific allergen levels were significantly correlated.
BACKGROUND: In this study we examine the variability among unstandardized cockroach allergen extracts. METHODS: We obtained 24 aqueous and glycerinated cockroach allergen extracts from nine manufacturers. We used previously characterized cockroach extracts, E2-Cg and E2-Ca, as references. The modified ninhydrin assay was used to determine protein concentration of each extract. Relative potencies of extracts were determined by competition ELISA, using a human allergic serum pool. Bla g 1 and Bla g 2 levels of glycerinatedGerman cockroach extracts were determined by ELISA using monoclonal antibodies. Extracts were also analysed by SDS-PAGE. RESULTS: Commercial cockroach allergen extracts had highly variable protein contents that were lower than the protein contents of the references. Electrophoretic data confirmed the presence of a variable number and intensity of protein bands in extracts among manufacturers. The relative potencies of the commercial extracts were between 10 and 782 BAU/mL for German cockroach and 10-250 BAU/mL for American cockroach. The mean Bla g 1 content of the commercial extracts was significantly lower than that of the reference (P = 0.001). The mean Bla g 2 content of the commercial extracts was higher than that of the E2-Cg reference but the Bla g 2 levels were more variable compared to Bla g 1. In glycerinatedGerman cockroach extracts, protein concentrations, relative potencies and specific allergen levels were significantly correlated (P < 0.001). CONCLUSION: Our tests indicate that commercially available cockroach allergen extracts are variable in protein content, electrophoretic banding patterns, relative potency and Bla g 2 levels. In glycerinatedGerman cockroach extracts, protein concentrations, relative potencies and specific allergen levels were significantly correlated.
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