Title: EXTRACTION OF PROTEIN FROM THE POLLEN OF COMMON RAGWEED, AMBROSIA ARTEMISIIFOLIA: USE OF THE METHOD FOR THE IMMUNOCHEMICAL QUANTIFICATION OF ANTIGEN A1/A2 IN POLLEN OF PLANTS GROWN IN CONTROLLED C02 ENVIRONMENTS.
Submitted to: Allergy and Immunology Meeting American Academy
Publication Type: Abstract Only
Publication Acceptance Date: March 15, 2002
Publication Date: N/A
Common ragweed pollen (Ambrosia artemisiifolia) is the major cause of allergic rhinitis (hay fever) in the United States. Methods for quantifying the antigenicity (allergenicity) of Ambrosia grown under varying levels of CO2 are of biochemical and ecological interest. We have developed a method for extracting pollen protein, which involves brief sonification of pollen suspended in buffer containing the mild non-ionic detergent Tween 20, followed by centrifugation. The described method requires about 30 minutes to process up to 20 pollen samples vs. 2-48 hours for the other methods. Total protein in the extract is highest using this method (7.3 mg/mL vs. 4.5-6.3 mg/mL). SDS-polyacrylamide gel electrophoresis demonstrated that the profile of proteins extracted by this method is similar to those obtained using the comparison methods. An enzyme-linked immunosorbent assay (ELISA) was used to determine antigen A1/A2 content in protein extracted from pollen of A. artemesiifolia plants grown under controlled [CO2]. Total pollen protein is elevated in plants grown at both elevated (600 ppm) and sub-ambient (280 ppm) [CO2] when compared with plants grown at standard CO2 conditions (370 ppm). Antigen A1/A2 as a fraction of total protein tends to be elevated in plants grown with elevated [CO2] when compared to ambient and sub-ambient conditions, though the differences are not significant. Ambrosia antigen A1/A2 levels (this study) as well as physiological and growth characteristics (previous studies) have been shown to be affected by rising atmospheric [CO2], suggesting that potentially significant health effects may result from global increases of this important greenhouse gas.