| pubmed-article:2461403 | pubmed:abstractText | Spore extracts from four common puffballs (Calvatia cyathiformis, Geaster saccatum, Pisolithus tinctorius, and Scleroderma areolatum) were fractionated by Sephadex G-75 gel filtration. Pooled fractions, analyzed by skin test or direct RAST, contained allergenic components over a molecular weight range of 10,500 to 20,000 daltons for all puffball spores studied except for S. areolatum (greater than 70,000 daltons). The Sephadex G-75 allergenic pool of C. cyathiformis was further fractionated by hydrophobic interaction chromatography. Fractions were successively eluted with 4 and 1 mol/L buffered saline solutions and water. The 1 mol/L fraction contained maximal allergenic activity as determined by RAST-inhibition studies. At least 15 Coomassie blue staining bands were identified after isoelectrofocusing, of which 7 were shown by immunoprinting to bind IgE from a serum pool of five patients with allergy. Our results demonstrate that a combination of gel filtration, hydrophobic interaction chromatography, RAST-inhibition, and immunoprinting studies can be used to fractionate, characterize, and standardize spore allergen extracts of puffballs for diagnostic and therapeutic purpose. | lld:pubmed |
