Detection of antibodies to proteases used in laundry detergents by the radioallergosorbent test.
Two proteases, Esperase and Alcalase, derived from Bacillus licheniformis and B. subtilis, respectively, are used in laundry products. In testing for the prevalence of IgE antibodies to these enzymes in sera among 300 laundry product workers, we experienced two problems in the establishment of a reliable RAST for these antigens. The first problem was the propensity of the allergen, Esperase, to undergo autolysis, suggesting that solid-phase Esperase might also lose reactivity through degradation. Treatment of Esperase with phenylmethylsulfonyl fluoride stabilized the enzyme and permitted the synthesis of a stable solid-phase antigen. The second problem was the finding that sera reactive with Esperase in the RAST were also reactive with Savinase, an enzyme from B. licheniformis to which the workers were not exposed. Immunochemical analyses of the three enzymes with specific rabbit antisera by gel diffusion and by two-site immunoradiometric assay demonstrated that they were not cross contaminated to any appreciable extent. RAST inhibition demonstrated that solid-phase Esperase possessed unique allergenic determinants in that the reactivity of IgE antibodies was inhibited by low concentrations of Esperase and only by very high concentrations of Alcalase and Savinase. In contrast, the reactivity of solid-phase Alcalase was occasionally inhibited equally well by Esperase and Alcalase. Most strikingly, the reaction of IgE antibodies with solid-phase Savinase was always inhibited by comparable quantities of Esperase, Alcalase, and Savinase. Thus, the establishment of the RAST for these proteases appears to require the use of phenylmethylsulfonyl fluoride to retard autolysis, and the results must be interpreted with caution because IgE antibodies in certain sera demonstrate cross-reactivity with Alcalase and Savinase.
D000906:Antibodies / Q000032:analysis*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D003902:Detergents / Q000493:pharmacokinetics*; D010450:Endopeptidases / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis; D009784:Occupational Diseases / Q000276:immunology*; D010447:Peptide Hydrolases / Q000276:immunology*; D011852:Radioallergosorbent Test; D012697:Serine Endopeptidases / Q000276:immunology; D013381:Subtilisins / Q000276:immunology; D013501:Surface-Active Agents / Q000493:pharmacokinetics*
D000906:Antibodies; D003902:Detergents; D013501:Surface-Active Agents; D007073:Immunoglobulin E; D010450:Endopeptidases; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; D013381:Subtilisins; C021553:mesentericopeptidase; C032923:microbial serine proteinases