Allergic potency of recombinant Fel d 1 is reduced by low concentrations of chlorine bleach

Background: Sodium hypochlorite (NaOCl), the primary component of household bleach, has been shown to alter the purified mouse allergen Mus m 1, such that antibody recognition, or immunogenicity, is lost. Results of initial experiments suggest that antibody recognition is lost at lower concentrations of NaOCl than those required to fragment Mus m 1. Objective: We sought to determine whether NaOCl had similar effects on recombinant (r)Fel d 1 and whether the loss of antibody recognition correlated with the loss of biologic activity, as measured with a basophil histamine release assay. Methods: Recombinant Fel d 1 was treated with increasing amounts of NaOCl, and the product of the reaction was analyzed by using SDS-PAGE, Western blotting, and ELISA. The biologic activity of NaOCl-treated rFel d 1 was analyzed with a basophil histamine release assay. Results: The protein fragmented at an NaOCl/rFel d 1 molar ratio of 7000, whereas cat-specific IgG recognition was lost at a lower molar ratio of 560. Basophil histamine release assays were performed to determine the effect of NaOCl on the biologic activity of rFel d 1. An NaOCl/protein molar ratio of 70 caused a significant reduction in histamine release from basophils of subjects with cat allergy. A molar ratio of 140 further inhibited histamine release by rFel d 1, suggesting a dose-response relationship between NaOCl and loss of biologic activity. Conclusions: NaOCl modifies rFel d 1, resulting in loss of immunogenicity and attenuation of biologic activity, as measured by its ability to stimulate basophil histamine release.