Analysis of human IgE epitope of Der f 2 with anti-Der f 2 mouse monoclonal antibodies.
Der f 2 is one of the major mite allergens recognized by human IgE antibodies of allergic patients. Using five anti-Der f 2 mouse monoclonal antibodies, human IgE epitopes of Der f 2 were analyzed. Among them, two monoclonal antibodies 15E11 and 13A4 inhibited the binding between Der f 2 and human IgE antibodies. To determine major IgE epitopes of Der f 2, epitopes for the monoclonal IgG antibodies were analyzed using 43 single site Der f 2 mutants constructed by site-directed mutagenesis. Binding ability of 13A4 and 15E11 was decreased by the amino acid replacement around the C-terminus, and around 73rd, respectively. These results suggest that the C-terminal portion and the central portion around 73rd of Der f 2 were recognized by human IgE antibodies as major epitopes. The location of the putative IgE epitopes on 3-D structure of Der f 2 is also discussed.
Nishiyama
C
C
Bioscience Research and Development Laboratory, Asahi Breweries Ltd, Ibaraki, Japan. chiharu.nishiyama@asahibeer.co.jp
Hatanaka
H
H
Ichikawa
S
S
Fukada
M
M
Akagawa-Chihara
M
M
Yuuki
T
T
Yokota
T
T
Inagaki
F
F
Okumura
Y
Y
D000485:Allergens / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D039741:Antigens, Dermatophagoides; D018604:Epitope Mapping; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology*; D007073:Immunoglobulin E / Q000276:immunology*; D051379:Mice; D008925:Mites; D008969:Molecular Sequence Data; D009154:Mutation
D000485:Allergens; D000911:Antibodies, Monoclonal; D000941:Antigens; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins; D016056:Immunodominant Epitopes; D007073:Immunoglobulin E