Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication
The authentication of meat products has become a global consumer concern in the food industry. Traditional methods such as PCR and mass spectrometry can identify and differentiate the source of meat, but they are time-consuming and require high-quality extracted DNA or protein for testing. As an alt...
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| Format: | Article |
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Elsevier Inc.
2024
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| author | Mohamad, Nornazliya Hashim, Amalia Mohd Khairil Mokhtar, Nur Fadhilah Yuswan, Mohd Hafis Mustafa, Shuhaimi |
| author_facet | Mohamad, Nornazliya Hashim, Amalia Mohd Khairil Mokhtar, Nur Fadhilah Yuswan, Mohd Hafis Mustafa, Shuhaimi |
| author_sort | Mohamad, Nornazliya |
| collection | UPM |
| description | The authentication of meat products has become a global consumer concern in the food industry. Traditional methods such as PCR and mass spectrometry can identify and differentiate the source of meat, but they are time-consuming and require high-quality extracted DNA or protein for testing. As an alternative, aptamer-based detection tools have been introduced, but their application in food authentication is still new. To date, there is a lack of data on the development of a porcine-specific aptamer that is specifically bound to a heat-stable protein. Hence, this study was conducted to screen, characterize and validate aptamers bound to any pork protein through SELEX process, combined with Next Generation Sequencing (NGS) and Liquid Chromatography Mass Spectrometry (LC-MS) analysis. The putative porcine-specific aptamers were selected after fourteen rounds of selection using centrifugal-ultrafiltration separation technique against five negative controls. The binding affinity test revealed that APT#A1 had the highest binding affinity with a dissociation constant of 27.61 ± 1.92 nM. However, the protein blotting results showed that the selected porcine-bound aptamers were not specific and could also bind to multiple proteins from negative samples. LC-MS analysis showed that the aptamers bound to troponin and tropomyosin subunits, and these proteins have potential as target markers for future authentication studies. Future research can focus on developing aptamers with higher specificity towards porcine protein and validating their feasibility as a practical tool for food authentication in real meat-based food samples. |
| first_indexed | 2024-04-09T03:51:40Z |
| format | Article |
| id | upm.eprints-105819 |
| institution | Universiti Putra Malaysia |
| last_indexed | 2024-04-09T03:51:40Z |
| publishDate | 2024 |
| publisher | Elsevier Inc. |
| record_format | dspace |
| spelling | upm.eprints-1058192024-04-05T03:30:59Z http://psasir.upm.edu.my/id/eprint/105819/ Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication Mohamad, Nornazliya Hashim, Amalia Mohd Khairil Mokhtar, Nur Fadhilah Yuswan, Mohd Hafis Mustafa, Shuhaimi The authentication of meat products has become a global consumer concern in the food industry. Traditional methods such as PCR and mass spectrometry can identify and differentiate the source of meat, but they are time-consuming and require high-quality extracted DNA or protein for testing. As an alternative, aptamer-based detection tools have been introduced, but their application in food authentication is still new. To date, there is a lack of data on the development of a porcine-specific aptamer that is specifically bound to a heat-stable protein. Hence, this study was conducted to screen, characterize and validate aptamers bound to any pork protein through SELEX process, combined with Next Generation Sequencing (NGS) and Liquid Chromatography Mass Spectrometry (LC-MS) analysis. The putative porcine-specific aptamers were selected after fourteen rounds of selection using centrifugal-ultrafiltration separation technique against five negative controls. The binding affinity test revealed that APT#A1 had the highest binding affinity with a dissociation constant of 27.61 ± 1.92 nM. However, the protein blotting results showed that the selected porcine-bound aptamers were not specific and could also bind to multiple proteins from negative samples. LC-MS analysis showed that the aptamers bound to troponin and tropomyosin subunits, and these proteins have potential as target markers for future authentication studies. Future research can focus on developing aptamers with higher specificity towards porcine protein and validating their feasibility as a practical tool for food authentication in real meat-based food samples. Elsevier Inc. 2024-01 Article PeerReviewed Mohamad, Nornazliya and Hashim, Amalia Mohd and Khairil Mokhtar, Nur Fadhilah and Yuswan, Mohd Hafis and Mustafa, Shuhaimi (2024) Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication. Microchemical Journal, 196. art. no. 109650. pp. 1-12. ISSN 0026-265X; ESSN: 1095-9149 https://www.sciencedirect.com/science/article/pii/S0026265X23012699 10.1016/j.microc.2023.109650 |
| spellingShingle | Mohamad, Nornazliya Hashim, Amalia Mohd Khairil Mokhtar, Nur Fadhilah Yuswan, Mohd Hafis Mustafa, Shuhaimi Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication |
| title | Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication |
| title_full | Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication |
| title_fullStr | Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication |
| title_full_unstemmed | Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication |
| title_short | Discovery of porcine proteins-binding DNA aptamer through SELEX and proteomics for pork authentication |
| title_sort | discovery of porcine proteins binding dna aptamer through selex and proteomics for pork authentication |
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