title: Purification strategy for recombinant Phl p 6 is applicable to the natural allergen and yields biochemically and immunologically comparable preparations.
abstract: The recombinant major grass pollen allergen Phl p 6 has been expressed with a N-terminal 6 x His-tag sequence and subsequently purified using nickel-chelating Sepharose. After cleavage of the tag-sequence, a second pass over the affinity chromatography revealed that even untagged rPhl p 6 bound tightly. In order to determine if that property is typical for Phl p 6, the natural allergen was purified in the same way starting with a grass pollen extract. Indeed, nPhl p 6 could be highly enriched in one step using nickel-chelating Sepharose. In addition to this new powerful purification method, the results provide further information in that the recombinant and natural allergens share a lot of properties, since biochemical characteristics are reflected in the purification strategies. The preparations of natural and recombinant Phl p 6 were used for comparative electrophoretic, chromatographic and immunological analysis which demonstrated high similarity.
authors.affiliation: Allergopharma Joachim Ganzer KG, Hermann-Körner-Strasse 52, 21465 Reinbek, Germany. roland.suck@allergopharma.de
authors.forename: Roland
authors.initials: R
authors.lastname: Suck
authors.forename: Bernhard
authors.initials: B
authors.lastname: Weber
authors.forename: Brigitte
authors.initials: B
authors.lastname: Schäffer
authors.forename: Ellen
authors.initials: E
authors.lastname: Diedrich
authors.forename: Timo
authors.initials: T
authors.lastname: Kamionka
authors.forename: Helmut
authors.initials: H
authors.lastname: Fiebig
authors.forename: Oliver
authors.initials: O
authors.lastname: Cromwell
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000485:Allergens; C095238:PHLPVI protein, Phleum pratense; D010940:Plant Proteins; D011994:Recombinant Proteins