High-throughput screening of T7 phage display and protein microarrays as a methodological approach for the identification of IgE-reactive components

Journal of Immunological Methods
Pablo San Segundo-AcostaRodrigo Barderas

Abstract

Olive pollen and yellow mustard seeds are major allergenic sources with high clinical relevance. To aid with the identification of IgE-reactive components, the development of sensitive methodological approaches is required. Here, we have combined T7 phage display and protein microarrays for the identification of allergenic peptides and mimotopes from olive pollen and mustard seeds. The identification of these allergenic sequences involved the construction and biopanning of T7 phage display libraries of mustard seeds and olive pollen using sera from allergic patients to both biological sources together with the construction of phage microarrays printed with 1536 monoclonal phages from the third/four rounds of biopanning. The screening of the phage microarrays with individual sera from allergic patients enabled the identification of 10 and 9 IgE-reactive unique amino acid sequences from olive pollen and mustard seeds, respectively. Five immunoreactive amino acid sequences displayed on phages were selected for their expression as His6-GST tag fusion proteins and validation. After immunological characterization, we assessed the IgE-reactivity of the constructs. Our results show that protein microarrays printed with T7 phages displa...Continue Reading

Citations

Jul 28, 2020·The Journal of Allergy and Clinical Immunology. in Practice·Robert G HamiltonJörg Kleine-Tebbe
May 29, 2019·Journal of Proteome Research·Pablo San Segundo-AcostaRodrigo Barderas
Oct 12, 2021·Journal of Proteome Research·Pablo San Segundo-AcostaRodrigo Barderas

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