Effects of Extraction Buffer on the Solubility and Immunoreactivity of the Pacific Oyster Allergens

Despite recent technological advances, novel allergenic protein discovery is limited by their low abundance, often due to specific physical characteristics restricting their recovery during the extraction process from various allergen sources. In this study, eight different extraction buffers were c...

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Bibliographic Details
Published inFoods Vol. 10; no. 2; p. 409
Main Authors Nugraha, Roni, Ruethers, Thimo, Johnston, Elecia B, Rolland, Jennifer M, O'Hehir, Robyn E, Kamath, Sandip D, Lopata, Andreas L
Format Journal Article
LanguageEnglish
Published Switzerland MDPI 12.02.2021
MDPI AG
Subjects
allergenomics
allergens
extraction buffer
immunoreactivity
proteomics
shellfish
immunoreactivity
Pacific oyster
allergenomics
shellfish
extraction buffer
proteomics
mollusk allergy
allergens
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Summary:Despite recent technological advances, novel allergenic protein discovery is limited by their low abundance, often due to specific physical characteristics restricting their recovery during the extraction process from various allergen sources. In this study, eight different extraction buffers were compared for their ability to recover proteins from Pacific oyster ( ). The protein composition was investigated using high resolution mass spectrometry. The antibody IgE-reactivity of each extract was determined using a pool of serum from five shellfish-allergic patients. Most of the investigated buffers showed good capacity to extract proteins from the Pacific oyster. In general, a higher concentration of proteins was recovered using high salt buffers or high pH buffers, subsequently revealing more IgE-reactive bands on immunoblotting. In contrast, low pH buffers resulted in a poor protein recovery and reduced IgE-reactivity. Discovery of additional IgE-reactive proteins in high salt buffers or high pH buffers was associated with an increase in allergen abundance in the extracts. In conclusion, increasing the ionic strength and pH of the buffer improves the solubility of allergenic proteins during the extraction process for oyster tissue. This strategy could also be applied for other difficult-to-extract allergen sources, thereby yielding an improved allergen panel for increased diagnostic efficiency.
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ISSN:2304-8158
2304-8158
DOI:10.3390/foods10020409