The invention relates to a method for quickly and effectively isolating and purifying five, namely the group 1, 2, 3, 10 and 13 allergens from grass pollen. The purification of said grass pollen is based on the inventive combination of hydrophobic interaction chromatography, gel filtration and cation exchange chromatography. The proteins obtained by the inventive method facilitate an improved diagnosis of pollen allergies and are used in pharmaceutical preparations for the therapy of pollenogenic diseases.