title: Rapid and efficient purification of Phleum pratense major allergens Phl p 1 and group Phl p 2/3 using a two-step procedure.
abstract: 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.
authors.affiliation: Allergopharma Joachim Ganzekg, Hermann-Körner-Strasse 52, 21465 Reinbek, Germany. allergopharmakg@csi.com
authors.forename: R
authors.initials: R
authors.lastname: Suck
authors.forename: S
authors.initials: S
authors.lastname: Hagen
authors.forename: O
authors.initials: O
authors.lastname: Cromwell
authors.forename: H
authors.initials: H
authors.lastname: Fiebig
mesh_terms: D000485:Allergens / Q000276:immunology / Q000302:isolation & purification*; D002850:Chromatography, Gel; D007073:Immunoglobulin E / Q000276:immunology; D007526:Isoelectric Point; D006109:Poaceae / Q000276:immunology*
chemical_list: D000485:Allergens; D007073:Immunoglobulin E