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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| Format: | Article |
| Language: | English |
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MDPI AG
2021-02-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/10/2/409 |
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| author | Roni Nugraha Thimo Ruethers Elecia B. Johnston Jennifer M. Rolland Robyn E. O’Hehir Sandip D. Kamath Andreas L. Lopata |
| author_facet | Roni Nugraha Thimo Ruethers Elecia B. Johnston Jennifer M. Rolland Robyn E. O’Hehir Sandip D. Kamath Andreas L. Lopata |
| author_sort | Roni Nugraha |
| collection | DOAJ |
| description | 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 (<i>Crassostrea gigas</i>). 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. |
| first_indexed | 2024-03-09T00:57:06Z |
| format | Article |
| id | doaj.art-ef2c5ab8633144a381add99ede560711 |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-09T00:57:06Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-ef2c5ab8633144a381add99ede5607112023-12-11T16:51:37ZengMDPI AGFoods2304-81582021-02-0110240910.3390/foods10020409Effects of Extraction Buffer on the Solubility and Immunoreactivity of the Pacific Oyster AllergensRoni Nugraha0Thimo Ruethers1Elecia B. Johnston2Jennifer M. Rolland3Robyn E. O’Hehir4Sandip D. Kamath5Andreas L. Lopata6Department of Aquatic Product Technology, Faculty of Fisheries and Marine Science, IPB University, Bogor 16680, West Java, IndonesiaMolecular Allergy Research Laboratory, College of Public Health, Medical and Veterinary Sciences, James Cook University, Douglas, QLD 4811, AustraliaMolecular Allergy Research Laboratory, College of Public Health, Medical and Veterinary Sciences, James Cook University, Douglas, QLD 4811, AustraliaThe Department of Immunology and Pathology, Monash University, Clayton, VIC 3800, AustraliaThe Department of Immunology and Pathology, Monash University, Clayton, VIC 3800, AustraliaMolecular Allergy Research Laboratory, College of Public Health, Medical and Veterinary Sciences, James Cook University, Douglas, QLD 4811, AustraliaMolecular Allergy Research Laboratory, College of Public Health, Medical and Veterinary Sciences, James Cook University, Douglas, QLD 4811, AustraliaDespite 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 (<i>Crassostrea gigas</i>). 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.https://www.mdpi.com/2304-8158/10/2/409allergensextraction bufferproteomicsallergenomicsshellfishimmunoreactivity |
| spellingShingle | Roni Nugraha Thimo Ruethers Elecia B. Johnston Jennifer M. Rolland Robyn E. O’Hehir Sandip D. Kamath Andreas L. Lopata Effects of Extraction Buffer on the Solubility and Immunoreactivity of the Pacific Oyster Allergens Foods allergens extraction buffer proteomics allergenomics shellfish immunoreactivity |
| title | Effects of Extraction Buffer on the Solubility and Immunoreactivity of the Pacific Oyster Allergens |
| title_full | Effects of Extraction Buffer on the Solubility and Immunoreactivity of the Pacific Oyster Allergens |
| title_fullStr | Effects of Extraction Buffer on the Solubility and Immunoreactivity of the Pacific Oyster Allergens |
| title_full_unstemmed | Effects of Extraction Buffer on the Solubility and Immunoreactivity of the Pacific Oyster Allergens |
| title_short | Effects of Extraction Buffer on the Solubility and Immunoreactivity of the Pacific Oyster Allergens |
| title_sort | effects of extraction buffer on the solubility and immunoreactivity of the pacific oyster allergens |
| topic | allergens extraction buffer proteomics allergenomics shellfish immunoreactivity |
| url | https://www.mdpi.com/2304-8158/10/2/409 |
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