Rapid and efficient purification of Phleum pratense major allergens Phl p 1 and group Phl p 2/3 using a two-step procedure.
The standardization of natural allergenic extracts and the characterization of recombinant allergens ensures a continuing requirement for highly purified natural allergens. The extraction and purification methods have to be reproducible and also preserve the biological and immunological activity of the allergen. A simple two-step purification system has been established in order to provide milligram amounts of purified natural Phl p 1 and Phl p 2/3. Both major allergens were separated from other proteins of timothy grass pollen extract in one step by hydrophobic interaction chromatography (HIC) under mild conditions. The allergens elute in the flow-through fraction while the rest of the proteins remain bound to the column. The very different molecular weights of Phl p 1 and Phl p 2/3 permitted separation of the allergens by a second step using gel filtration.
Suck
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Allergopharma Joachim Ganzekg, Hermann-Körner-Strasse 52, 21465 Reinbek, Germany. allergopharmakg@csi.com
Hagen
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S
Cromwell
O
O
Fiebig
H
H
D000485:Allergens / Q000276:immunology / Q000302:isolation & purification*; D002850:Chromatography, Gel; D007073:Immunoglobulin E / Q000276:immunology; D007526:Isoelectric Point; D006109:Poaceae / Q000276:immunology*
D000485:Allergens; D007073:Immunoglobulin E