Isolation and partial characterization of three allergens of timothy pollen

Three allergens, antigen Nos. 3, 25, and 30, were isolated from freeze-dried aqueous extract of timothy pollen by various combinations of anionic and cationic exchange and gel chromatography. The allergens were all of protein nature and contained less than 2% (w/w) immunochem;cally detectable impurities. Molecular weights and pI's were determined to less than 10 X 10(3), 15 X 10(3) and 3.9, 4.5, 9.4, respectively, and antigen 25 was determined as the major component of the earlier isolated antigen B. amino acid analyses performed an antigens 3 and 30 revealed large variation s in the amino acid composition. The allergenic activities were verified by means of RAST inhibition and pick tests and proved antigen 25 to be quantitatively the most important allergen of timothy pollen.