Comparison of three horizontal two-dimensional electrophoretic techniques to separate grass pollen allergens.

The identification and characterization of allergenic components are important for improving both diagnosis and therapy of allergy. We have studied grass pollen crude extract for better characterization of the allergen repertoire recognized by allergic patient IgE antibodies. Two-dimensional electrophoresis are the methods of choice to define physico-chemical characteristics of the allergens as they give isoelectric points and molecular mass of the analyzed samples. We compared the advantages and the drawbacks of three different ways to perform a two-dimensional electrophoresis. First, two gel media were compared to run native isoelectric focusing (IEF) with carrier ampholytes: agarose and acrylamide. With the introduction of immobilized pH gradient, highly resolutive and reproducible 2D electrophoresis could be obtained. This technique was also applied to separate the allergens of Dactylis glomerata pollen.