Taruna Khurana1, Maggie Collison1, Fook Tim Chew2, Jay E Slater3. 1. Laboratory of Immunobiochemistry, Division of Bacterial, Parasitic, and Allergenic Products, US Food and Drug Administration, Bethesda, Maryland. 2. Allergy and Molecular Immunology Laboratory, Department of Biological Sciences, National University of Singapore, Singapore. 3. Laboratory of Immunobiochemistry, Division of Bacterial, Parasitic, and Allergenic Products, US Food and Drug Administration, Bethesda, Maryland. Electronic address: jay.slater@fda.hhs.gov.
Abstract
BACKGROUND: The IgE response to cockroach allergens is thought to be associated with asthma. German cockroach (GCr) allergen extract is a complex mixture of allergens, and the identification and characterization of immunodominant allergens is important for the effective diagnosis and treatment of GCr-induced asthma. OBJECTIVE: To characterize a novel GCr allergen homologous to the American cockroach allergen Per a 3. METHODS: GCr-specific avian monoclonal antibodies were used for direct immunoprecipitation of specific targets from whole-body GCr extract. Precipitated protein was identified by mass spectrometry and sequence analysis. Putative recombinant protein also was expressed, purified, and used for determination of allergenicity, determined by IgE enzyme-linked immunosorbent assay with serum from 61 GCr-allergic patients. The identified target also was analyzed for heat stability using a bead-based assay. RESULTS: The immunoprecipitated target of monoclonal antibody 2A1 was identified as a novel allergen of GCr homologous to American cockroach allergen Per a 3. This homolog, designated Bla g 3, has an apparent mass of 78 kDa, can be measured in GCr extract using antibody 2A1, and is a heat-stable protein. Screening of 61 serum samples from GCr-allergic patients showed a 22% prevalence of Bla g 3-specific IgE. CONCLUSION: Bla g 3 is a GCr allergen with structural homology to American cockroach allergen Per a 3.
BACKGROUND: The IgE response to cockroach allergens is thought to be associated with asthma. German cockroach (GCr) allergen extract is a complex mixture of allergens, and the identification and characterization of immunodominant allergens is important for the effective diagnosis and treatment of GCr-induced asthma. OBJECTIVE: To characterize a novel GCr allergen homologous to the American cockroach allergen Per a 3. METHODS:GCr-specific avian monoclonal antibodies were used for direct immunoprecipitation of specific targets from whole-body GCr extract. Precipitated protein was identified by mass spectrometry and sequence analysis. Putative recombinant protein also was expressed, purified, and used for determination of allergenicity, determined by IgE enzyme-linked immunosorbent assay with serum from 61 GCr-allergicpatients. The identified target also was analyzed for heat stability using a bead-based assay. RESULTS: The immunoprecipitated target of monoclonal antibody 2A1 was identified as a novel allergen of GCr homologous to American cockroach allergen Per a 3. This homolog, designated Bla g 3, has an apparent mass of 78 kDa, can be measured in GCr extract using antibody 2A1, and is a heat-stable protein. Screening of 61 serum samples from GCr-allergicpatients showed a 22% prevalence of Bla g 3-specific IgE. CONCLUSION: Bla g 3 is a GCr allergen with structural homology to American cockroach allergen Per a 3.
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