Identifying the stability of a new wheat gliadin extract by protein analysis, skin tests and cell degranulation assay
Issued Date
2025-12-01
Resource Type
ISSN
0125877X
eISSN
22288694
Scopus ID
2-s2.0-105029372139
Pubmed ID
37578480
Journal Title
Asian Pacific Journal of Allergy and Immunology
Volume
43
Issue
4
Start Page
856
End Page
863
Rights Holder(s)
SCOPUS
Bibliographic Citation
Asian Pacific Journal of Allergy and Immunology Vol.43 No.4 (2025) , 856-863
Suggested Citation
Chansangsawat C., Piboonpocanun S., Ubonsri P., Pacharn P., Srisuwatchari W., Kanchanapoomi K., Visitsunthorn N., Jirapongsananuruk O. Identifying the stability of a new wheat gliadin extract by protein analysis, skin tests and cell degranulation assay. Asian Pacific Journal of Allergy and Immunology Vol.43 No.4 (2025) , 856-863. 863. doi:10.12932/ap-010323-1553 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/115043
Title
Identifying the stability of a new wheat gliadin extract by protein analysis, skin tests and cell degranulation assay
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Abstract
Background: The commercial wheat extract for skin prick test (SPT) provides less sensitivity to predict wheat allergy, compared to in-house gliadin extracts. SPT is a preferred method to study extract stability as it is the aim of developing extract. The role of cell degranulation assay, a functional assay with the same mechanism as SPT, is not widely used to determine extract stability. Objective: To study the stability of in-house gliadin extracts stored at different periods, by using protein analysis, SPT and degranulation assay of humanized rat basophilic-leukemia (RBL-SX38) cells. Methods: Patients with a history of wheat allergy and positive SPT to wheat, were recruited. The gliadin extracts stored for 1, 6, 9, and 12 months at 2–8°C were used in SDS-PAGE, SPT and cell degranulation assay. The cell degranulation was determined by β-hexosaminidase release. Results: Forty children were recruited. The gliadin extract stored for 9 and 12 months provided lighter protein bands than 1 and 6 months. However, the wheal diameters from SPT using extracts stored at different periods, were not significantly different (p = 0.09). There were also no significant differences of the β-hexosaminidase released using 0.1 and 1 μg/mL of gliadin extracts stored at different periods (p > 0.05). The 10 μg/mL of gliadin extracts stored at longer periods, significantly stimulated higher β-hexosaminidase release (p = 0.01). The extracts were sterile at all storage times. Conclusions: To determine the stability of in-house gliadin extracts, SPT or cell degranulation assay provided additional information to SDS-PAGE. The extracts were stable for up to 12 months.