Effects of extraction buffer on the solubility and immunoreactivity of the pacific oyster allergens

Nugraha, Roni, Ruethers, Thimo, Johnston, Elecia B., Rolland, Jennifer M., O’Hehir, Robyn E., Kamath, Sandip D., and Lopata, Andreas L. (2021) Effects of extraction buffer on the solubility and immunoreactivity of the pacific oyster allergens. Foods, 10 (2). 409.

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Abstract

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 (Crassostrea gigas). 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.

Item ID: 70531
Item Type: Article (Research - C1)
ISSN: 2304-8158
Keywords: Allergenomics, Allergens, Extraction buffer, Immunoreactivity, Mollusk allergy, Pacific oyster, Proteomics, Shellfish
Copyright Information: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license https://creativecommons.org/licenses/by/4.0/
Date Deposited: 15 Mar 2022 00:04
FoR Codes: 30 AGRICULTURAL, VETERINARY AND FOOD SCIENCES > 3006 Food sciences > 300605 Food safety, traceability, certification and authenticity @ 20%
32 BIOMEDICAL AND CLINICAL SCIENCES > 3204 Immunology > 320401 Allergy @ 60%
32 BIOMEDICAL AND CLINICAL SCIENCES > 3206 Medical biotechnology > 320602 Medical biotechnology diagnostics (incl. biosensors) @ 20%
SEO Codes: 20 HEALTH > 2001 Clinical health > 200101 Diagnosis of human diseases and conditions @ 50%
28 EXPANDING KNOWLEDGE > 2801 Expanding knowledge > 280101 Expanding knowledge in the agricultural, food and veterinary sciences @ 30%
10 ANIMAL PRODUCTION AND ANIMAL PRIMARY PRODUCTS > 1002 Fisheries - aquaculture > 100203 Aquaculture molluscs (excl. oysters) @ 20%
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