Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis
An optimised protocol for the extraction of allergenic proteins in fish was developed. Twelve existing or modified extraction buffers were evaluated based on the amount and quality of obtained 12 kDa IgG-binding protein (major fish allergen parvalbumin) by SDS-PAGE, immunoblotting, and ELISA. The re...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Czech Academy of Agricultural Sciences
2017-02-01
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| Series: | Czech Journal of Food Sciences |
| Subjects: | |
| Online Access: | https://cjfs.agriculturejournals.cz/artkey/cjf-201701-0005_optimisation-of-an-extraction-technique-of-fish-allergens-suitable-for-detection-and-diagnosis.php |
| _version_ | 1797899590221955072 |
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| author | Jiaju Ma Tushar Ramesh Pavase Zhen-Xing Li Hong Lin |
| author_facet | Jiaju Ma Tushar Ramesh Pavase Zhen-Xing Li Hong Lin |
| author_sort | Jiaju Ma |
| collection | DOAJ |
| description | An optimised protocol for the extraction of allergenic proteins in fish was developed. Twelve existing or modified extraction buffers were evaluated based on the amount and quality of obtained 12 kDa IgG-binding protein (major fish allergen parvalbumin) by SDS-PAGE, immunoblotting, and ELISA. The results indicated that using DL-Dithiothreitol during the extraction process, the stability and functionality of the final extract were significantly increased. Solution 3 (containing Tris-HCl, Glycine, and DL-Dithiothreitol) yielded the highest amount of parvalbumin. The extract also induced a higher allergenic reactivity to the three tested human sera by immunoblotting. The results indicated that solution 3 provided better results in fish allergen detection and diagnosis. In conclusion, our study discovered and established solution 3 (Tris-HCl + Glycine + DTT) as the most potential buffer to overcome the conventional allergen extraction buffer and therefore it is highly recommended as a substitute optimised extraction buffer. |
| first_indexed | 2024-04-10T08:32:24Z |
| format | Article |
| id | doaj.art-203f6453a9f34fb58170627a1e053a27 |
| institution | Directory Open Access Journal |
| issn | 1212-1800 1805-9317 |
| language | English |
| last_indexed | 2024-04-10T08:32:24Z |
| publishDate | 2017-02-01 |
| publisher | Czech Academy of Agricultural Sciences |
| record_format | Article |
| series | Czech Journal of Food Sciences |
| spelling | doaj.art-203f6453a9f34fb58170627a1e053a272023-02-23T03:28:25ZengCzech Academy of Agricultural SciencesCzech Journal of Food Sciences1212-18001805-93172017-02-01351243110.17221/578/2015-CJFScjf-201701-0005Optimisation of an extraction technique of fish allergens suitable for detection and diagnosisJiaju Ma0Tushar Ramesh Pavase1Zhen-Xing LiHong Lin2Food Safety Laboratory, Ocean University of China, Qingdao, P.R. ChinaFood Safety Laboratory, Ocean University of China, Qingdao, P.R. ChinaFood Safety Laboratory, Ocean University of China, Qingdao, P.R. ChinaAn optimised protocol for the extraction of allergenic proteins in fish was developed. Twelve existing or modified extraction buffers were evaluated based on the amount and quality of obtained 12 kDa IgG-binding protein (major fish allergen parvalbumin) by SDS-PAGE, immunoblotting, and ELISA. The results indicated that using DL-Dithiothreitol during the extraction process, the stability and functionality of the final extract were significantly increased. Solution 3 (containing Tris-HCl, Glycine, and DL-Dithiothreitol) yielded the highest amount of parvalbumin. The extract also induced a higher allergenic reactivity to the three tested human sera by immunoblotting. The results indicated that solution 3 provided better results in fish allergen detection and diagnosis. In conclusion, our study discovered and established solution 3 (Tris-HCl + Glycine + DTT) as the most potential buffer to overcome the conventional allergen extraction buffer and therefore it is highly recommended as a substitute optimised extraction buffer.https://cjfs.agriculturejournals.cz/artkey/cjf-201701-0005_optimisation-of-an-extraction-technique-of-fish-allergens-suitable-for-detection-and-diagnosis.phpparvalbuminpurificationelisaimmunoblotting |
| spellingShingle | Jiaju Ma Tushar Ramesh Pavase Zhen-Xing Li Hong Lin Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis Czech Journal of Food Sciences parvalbumin purification elisa immunoblotting |
| title | Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis |
| title_full | Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis |
| title_fullStr | Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis |
| title_full_unstemmed | Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis |
| title_short | Optimisation of an extraction technique of fish allergens suitable for detection and diagnosis |
| title_sort | optimisation of an extraction technique of fish allergens suitable for detection and diagnosis |
| topic | parvalbumin purification elisa immunoblotting |
| url | https://cjfs.agriculturejournals.cz/artkey/cjf-201701-0005_optimisation-of-an-extraction-technique-of-fish-allergens-suitable-for-detection-and-diagnosis.php |
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