Development of peptide arrays for detection of IgE-binding epitopes in cow's milk allergens.
Peptide arrays have become versatile tools for high throughput screening assays in biomedical and pharmaceutical research. In this study, we constructed a peptide array that contained linear peptide fragments reported as IgE-binding epitopes for cow's milk allergy (CMA). Various peptides with different solubility in aqueous solutions were dissolved in the buffer solutions containing sodium dodecyl sulfate, and we achieved a consistent spotting of peptide solutions using a piezoelectric ceramic micropump. The IgE-binding patterns were successfully detected by observing the binding of Alexa 647-labeled anti-human IgE using sera from CMA patients. Our technique in this study will provide a potent capability for the development of a peptide array for mapping IgE-epitopes in milk proteins, and it will help researchers better understand the IgE-epitopes associated with the clinical outcome of CMA.
Matsumoto
Naoki
N
MEXT Innovative Research Center for Preventive Medical Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
Okochi
Mina
M
Matsushima
Miyoko
M
Ogawa
Akiko
A
Takase
Tomokazu
T
Yoshida
Yasuko
Y
Kawase
Mitsuo
M
Isobe
Ken-Ichi
K
Kawabe
Tsutomu
T
Honda
Hiroyuki
H
D000485:Allergens / Q000276:immunology*; D000818:Animals; D002417:Cattle; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology*; D016269:Milk Hypersensitivity / Q000276:immunology*; D008894:Milk Proteins / Q000276:immunology*; D010455:Peptides / Q000276:immunology*; D040081:Protein Array Analysis; D011485:Protein Binding
D000485:Allergens; D000939:Epitopes; D008894:Milk Proteins; D010455:Peptides; D007073:Immunoglobulin E