title: Affinity purification of a major Alternaria allergen using a monoclonal antibody.
abstract: An extract of Alternaria (ALT) was passed through an affinity column containing a monoclonal antibody directed to ALT. After washing the column, a single glycoprotein was eluted using 0.1 M glycine (pH 3.0). This glycoprotein accounted for 13% of the dry weight of our ALT preparation and had a carbohydrate to protein ratio of 0.35 as compared with 3.2 for the whole ALT extract. Its molecular weight was 70 kD by SDS-PAGE and isoelectric point was 3.5 by IEF. Though individual sensitivities varied, 14/16 patients skin reactive to ALT were also reactive to this glycoprotein. Quantitative skin tests of extracts adjusted to the same dry weight per volume showed that it required about a four times greater concentration of the purified glycoprotein to give a wheal size equal to whole Alternaria. The ability to purify large amounts of this allergen with affinity chromatography should make its complete characterization possible.
authors.affiliation: Children's Mercy Hospital, University of Health Sciences, Kansas City, Missouri.
authors.forename: J
authors.initials: J
authors.lastname: Portnoy
authors.forename: I
authors.initials: I
authors.lastname: Olson
authors.forename: F
authors.initials: F
authors.lastname: Pacheco
authors.forename: C
authors.initials: C
authors.lastname: Barnes
mesh_terms: D000293:Adolescent; D000485:Allergens / Q000302:isolation & purification*; D000528:Alternaria / Q000276:immunology*; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal / Q000302:isolation & purification*; D002648:Child; D002675:Child, Preschool; D002846:Chromatography, Affinity / Q000379:methods; D006801:Humans; D003904:Mitosporic Fungi / Q000276:immunology*
chemical_list: D000485:Allergens; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal