HPLC purification of the main allergen of Parietaria judaica pollen.
A two-step purification procedure of Par j I from the whole Parietaria judaica pollen extract is described. The first step consisted of gel filtration HPLC using a TSKG 3000 SW column, and 0.1% trifluoroacetic acid as the eluant. By this method, proteins were separated from the highly colored material present in the extract. Then, Par j I-containing fractions were chromatographed on a reversed-phase HPLC column (Vydac C4) using an acetonitrile gradient. This second step yielded pure Par j I as assessed by SDS-PAGE and CIE. Previously reported microheterogeneity was still observed, but amino acid analysis of various RP-HPLC fractions suggested that the heterogeneity of Par j I might not be due to changes in its primary structure. Allergenic activity of Par j I was shown to be retained after the purification procedure by several immunochemical techniques.
Polo
F
F
Departamento Investigación, Alergia e Inmunología Abelló S.A., Madrid, Spain.
Ayuso
R
R
Carreira
J
J
D000485:Allergens / Q000032:analysis / Q000302:isolation & purification*; D000596:Amino Acids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D010940:Plant Proteins; D011058:Pollen / Q000032:analysis / Q000276:immunology*
D000485:Allergens; D000596:Amino Acids; C083860:Par j I protein, Parietaria judaica; D010940:Plant Proteins