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Characterization of 4 Cupressus arizonica extracts using different solvents
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Characterization Of 4 Cupressus arizonica Extracts Using Different Solvents
J. Carnés1, J. Carnés Isidro2, A. Ferrer3, J. Sastre4, C. Andreu3, E. Fernández-Caldas1; 1C.B.F. LETI S.A., Tres Cantos, SPAIN, 2Hospital Central de la Defensa, Madrid, SPAIN, 3Hospital de la Vega Baja, Alicante, SPAIN, 4Fundación Jiménez Díaz, Madrid, SPAIN. RATIONALE: Cypress spp. pollen induced pollinosis is increasing worldwide. The objective of this study was to characterize 4 Cupressus arizonica extracts using different solvents and in vivo and in vitro techniques. METHODS: Commercially available Cupressus arizonica pollen was used to prepare 4 types of extracts extracts. Extract A was extracted in PBS (0.01M); extract B in ammonium bicarbonate (40 mM); extract C, in ammonium bicarbonate (40 mM), followed by precipitation with ammonium sulfate; and extract D consisted of the supernatant obtained after the centrifugation of extract C. All extracts were characterized by SDS-PAGE and immunoblot. Cup a 1 content was measured by scanning densitometry. Thirty individuals were skin tested with the 4 extracts at 2 mg/ml of freeze-dried material. Specific IgE was evaluated by direct ELISA and immunoblots. Polysaccharide content was evaluated by the Anthrone method. RESULTS: The diagnosis capacity of extract C was better than of extracts A, B and D. All skin prick tests were negative with extract D. Extract C had the highest protein and major allergen content and induced greater wheal sizes than the other extracts (p<0.05). Cup a 1 content was 7.2, 14.1 and 23.1 µg/mg of freeze-dried material in extracts A, B and C, respectively. No differences were found in the polysaccharide content in these extracts. CONCLUSIONS: The increase in the protein and major allergen content and the removal of irrelevant material produces a better extract for the in vivo and in vitro diagnosis of C. arizonica induced respiratory allergies. Funding: Self-funded
Abstracts S139
SUNDAY
J ALLERGY CLIN IMMUNOL VOLUME 113, NUMBER 2
Characterization Of 4 Cupressus arizonica Extracts Using Different Solvents
J. Carnés1, J. Carnés Isidro2, A. Ferrer3, J. Sastre4, C. Andreu3, E. Fernández-Caldas1; 1C.B.F. LETI S.A., Tres Cantos, SPAIN, 2Hospital Central de la Defensa, Madrid, SPAIN, 3Hospital de la Vega Baja, Alicante, SPAIN, 4Fundación Jiménez Díaz, Madrid, SPAIN. RATIONALE: Cypress spp. pollen induced pollinosis is increasing worldwide. The objective of this study was to characterize 4 Cupressus arizonica extracts using different solvents and in vivo and in vitro techniques. METHODS: Commercially available Cupressus arizonica pollen was used to prepare 4 types of extracts extracts. Extract A was extracted in PBS (0.01M); extract B in ammonium bicarbonate (40 mM); extract C, in ammonium bicarbonate (40 mM), followed by precipitation with ammonium sulfate; and extract D consisted of the supernatant obtained after the centrifugation of extract C. All extracts were characterized by SDS-PAGE and immunoblot. Cup a 1 content was measured by scanning densitometry. Thirty individuals were skin tested with the 4 extracts at 2 mg/ml of freeze-dried material. Specific IgE was evaluated by direct ELISA and immunoblots. Polysaccharide content was evaluated by the Anthrone method. RESULTS: The diagnosis capacity of extract C was better than of extracts A, B and D. All skin prick tests were negative with extract D. Extract C had the highest protein and major allergen content and induced greater wheal sizes than the other extracts (p<0.05). Cup a 1 content was 7.2, 14.1 and 23.1 µg/mg of freeze-dried material in extracts A, B and C, respectively. No differences were found in the polysaccharide content in these extracts. CONCLUSIONS: The increase in the protein and major allergen content and the removal of irrelevant material produces a better extract for the in vivo and in vitro diagnosis of C. arizonica induced respiratory allergies. Funding: Self-funded
Abstracts S139
SUNDAY
J ALLERGY CLIN IMMUNOL VOLUME 113, NUMBER 2