Purification of a basic glycoprotein allergen from pollen of timothy by high-performance liquid chromatography.
The use of high-performance ion-exchange and size-exclusion chromatography in the purification of the basic timothy pollen allergen antigen 30 (Ag 30) was investigated. The most efficient purification was achieved when an initial purification step on a CM-Sepharose CL-6B column was followed by chromatography on Mono S and TSK G 2000 SW columns. This procedure was highly reproducible and well suited for semi-preparative scale purification of the allergen. The purified allergen gave one band on isoelectric focusing, corresponding to a pI of 9.30. On fused rocket immunoelectrophoresis a single precipitate was obtained that coincided with the allergenic activity.
D000485:Allergens / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D006023:Glycoproteins / Q000302:isolation & purification*; D007122:Immunoelectrophoresis; D007525:Isoelectric Focusing; D008970:Molecular Weight; D006109:Poaceae / Q000032:analysis*; D011058:Pollen / Q000032:analysis*
D000485:Allergens; D006023:Glycoproteins