Production of Recombinant Allergens Phl p 1 and Amb a 1 for Detection of Class E Immunoglobulins

Abstract

Recombinant major allergens Phl p 1 from meadow timothy (Phleum pratense) and Amb a 1 from ragweed (Ambrosia artemisiifolia) were obtained in E. coli cells. Allergen purification procedures were developed using anion exchange and affinity chromatography, from which we obtain 10 mg Phl p 1 and 7.5 mg Amb a 1 from 1 L of culture medium with an electrophoretic purity of at least 90%. To analyze the ability of IgE antibodies to bind Phl p 1 and Amb a 1, sera obtained from people with IgE to various respiratory allergens, identified by the RIDA method, were used. Part of the sera bound Phl p 1 or Amb a 1, which confirmed the immunoreactivity of the obtained recombinant proteins. Antibody titers to Amb a 1 correlated with RIDA data on sensitivity to a mixture of herbs and rye pollen. Antibody titers to Phl p 1 did not correlate with RIDA data. Of the 20 positive sera selected, 73 and 89% of the sera recognizing Phl p 1 or Amb a 1, respectively, had antibodies to the cross-reactive alder/birch/hazel allergen group.