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  • Title: Sequential epitopes of Dermatophagoides farinae allergens identified using peptide microarray-based immunoassay.
    Author: Cui Y, Teng F, Yu L, Zhou Y, Wang N, Zhang C, Yang L.
    Journal: IUBMB Life; 2016 Oct; 68(10):792-8. PubMed ID: 27481284.
    Abstract:
    House dust mites produce over 30 proteins proposed to induce immunoglobulin E (IgE) antibody production in patients. Continued identification of IgE-binding epitopes of these allergens is critical to advancing diagnosis and treatment of allergic disease. To identify possible sequential IgE-binding epitopes of the major- and mid-potency allergens from the house dust mite Dermatophagoides farinae by peptide microarray-based immunoassay, nucleotide sequences of D. farinae allergens (Der f) 1, 2, 4, 5, and 7 were used to generate overlapping peptides covering the full protein sequences minus signal peptides. Short peptides were printed onto microarray chips. Because asthma occurs as a symptom of mite allergy more commonly among children than adults, the peptide chips were exposed to sera pooled from six serum-positive pediatric patients with D. farinae hypersensitivity and six serum-negative control children for screening sequential IgE-binding epitopes by IgE immunolabeling. Higher-than-average immunolabel signal intensity was observed for 21 short peptides in the serum-positive group (P < 0.01). Due to sequence overlap, these 21 signals represented four fragments of Der f 1 (amino acid positions 46-53, 71-78, 99-110, 179-186), three fragments of Der f 2 (15-22, 80-89, 106-113), six fragments of Der f 4 (69-82, 107-116, 225-232, 261-268, 355-365, 483-496), one fragment of Der f 5 (102-109), and three fragments of Der f 7 (32-39, 52-64, 100-107). These findings not only demonstrate the utility of a peptide microarray immunoassay in identifying epitopes for these allergens, but also provide a foundation for future exploration of specific immunotherapies. © 2016 IUBMB Life, 68(10):792-798, 2016.
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