Identification of the IgE-binding epitopes in main dust mite allergen Der p 1.
- Author:
Zhi-mei ZHONG
1
;
Chuan-dong ZHENG
;
Fang WANG
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Animals; Antigens, Dermatophagoides; immunology; Arthropod Proteins; immunology; Binding Sites, Antibody; Cysteine Endopeptidases; immunology; Epitopes; immunology; Immunoglobulin E; immunology; Lymphokines; immunology; Mites; immunology; Molecular Sequence Data
- From: Journal of Southern Medical University 2011;31(7):1183-1186
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify the IgE-binding epitopes in the allergen Der p 1 of main house dust mites, which can be recognized by the specific IgE in the sera from allergic individuals, and obtain a hypoallergen derived from the T-B epitope fused peptide for potential use in specific immunotherapy (SIT).
METHODSThirty-one peptides containing 15 amino acids each, which covered the full 222 amino acids of Der p 1 protein sequence, were synthesized on the cellulous membrane by solid-phase peptide (SPOTs) synthesis, with 8 overlapping amino acids between every two neighboring peptides. The membrane bearing the spots of the synthesized peptides were incubated with the allergic serum pools consisting of the sera from 5 allergic individuals. The membrane was then probed with HRP-conjugated anti-human IgE, followed by enhanced chemiluminescence (ECL) for visualization and gray scale analysis of the positive peptide spots.
RESULTSThree strong IgE-binding epitopes were identified in the amino acid sequence of Der p 1 molecule, namely Ep1 (amino acids 85-99), Ep2 (amino acids 106-120) and Ep3 (amino acids 190-204).
CONCLUSIONThe 3 IgE-binding epitopes (B cell epitopes) identified in Der p 1 confirm the presence of linear epitopes in Der p 1, suggesting the possibility of constructing T/B epitope-fused hypoallergens.