A novel variable antibody fragment dimerized by the dHLX peptide with enhanced affinity against amantadine compared to its corresponding scFv antibody
Amantadine (AMA) is an illegally used antiviral drug in the poultry industry, it is necessary to establish a fast, accurate and time-saving detection method for poultry food. The antibody-based immunoassay can achieve fast and accurate requirements. We developed a recombinant antibody-based specific...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2018-01-01
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| Series: | Food and Agricultural Immunology |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/09540105.2017.1368459 |
| _version_ | 1818694741448982528 |
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| author | Sanlei Xie Kai Wen Tao Peng Jianyi Wang Kai Yao Haiyang Jiang |
| author_facet | Sanlei Xie Kai Wen Tao Peng Jianyi Wang Kai Yao Haiyang Jiang |
| author_sort | Sanlei Xie |
| collection | DOAJ |
| description | Amantadine (AMA) is an illegally used antiviral drug in the poultry industry, it is necessary to establish a fast, accurate and time-saving detection method for poultry food. The antibody-based immunoassay can achieve fast and accurate requirements. We developed a recombinant antibody-based specificity immunoassay for AMA. In the recombinant antibody, the heavy chain variable region (VH) is connected covalently with the light chain variable region (VL) by the artificial linker. Here, two recombinant antibodies’ single-chain variable fragment (scFv) and scFv-dHLX were constructed and functionally expressed in the periplasm of Escherichia coli. The helix-turn-helix peptide was utilized to dimerize VH and VL similar to the IgG counterpart. The ScFv-dHLX protein showed a higher binding ability and affinity resulting in improvement of in vitro affinity activity over its corresponding scFv. Our results not only indicated scFv-dHLX as an alternative for scFv in analytical application, but also offered a novel and efficient hetero-dimerization pattern of VH and VL leading to enhanced affinity. |
| first_indexed | 2024-12-17T13:34:24Z |
| format | Article |
| id | doaj.art-0b2574ae36e6435494a4c957750f213d |
| institution | Directory Open Access Journal |
| issn | 0954-0105 1465-3443 |
| language | English |
| last_indexed | 2024-12-17T13:34:24Z |
| publishDate | 2018-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Food and Agricultural Immunology |
| spelling | doaj.art-0b2574ae36e6435494a4c957750f213d2022-12-21T21:46:28ZengTaylor & Francis GroupFood and Agricultural Immunology0954-01051465-34432018-01-0129124425310.1080/09540105.2017.13684591368459A novel variable antibody fragment dimerized by the dHLX peptide with enhanced affinity against amantadine compared to its corresponding scFv antibodySanlei Xie0Kai Wen1Tao Peng2Jianyi Wang3Kai Yao4Haiyang Jiang5Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityBeijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityBeijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityBeijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityBeijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityBeijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityAmantadine (AMA) is an illegally used antiviral drug in the poultry industry, it is necessary to establish a fast, accurate and time-saving detection method for poultry food. The antibody-based immunoassay can achieve fast and accurate requirements. We developed a recombinant antibody-based specificity immunoassay for AMA. In the recombinant antibody, the heavy chain variable region (VH) is connected covalently with the light chain variable region (VL) by the artificial linker. Here, two recombinant antibodies’ single-chain variable fragment (scFv) and scFv-dHLX were constructed and functionally expressed in the periplasm of Escherichia coli. The helix-turn-helix peptide was utilized to dimerize VH and VL similar to the IgG counterpart. The ScFv-dHLX protein showed a higher binding ability and affinity resulting in improvement of in vitro affinity activity over its corresponding scFv. Our results not only indicated scFv-dHLX as an alternative for scFv in analytical application, but also offered a novel and efficient hetero-dimerization pattern of VH and VL leading to enhanced affinity.http://dx.doi.org/10.1080/09540105.2017.1368459amantadineaffinitybinding abilityscfvscfv-dhlx |
| spellingShingle | Sanlei Xie Kai Wen Tao Peng Jianyi Wang Kai Yao Haiyang Jiang A novel variable antibody fragment dimerized by the dHLX peptide with enhanced affinity against amantadine compared to its corresponding scFv antibody Food and Agricultural Immunology amantadine affinity binding ability scfv scfv-dhlx |
| title | A novel variable antibody fragment dimerized by the dHLX peptide with enhanced affinity against amantadine compared to its corresponding scFv antibody |
| title_full | A novel variable antibody fragment dimerized by the dHLX peptide with enhanced affinity against amantadine compared to its corresponding scFv antibody |
| title_fullStr | A novel variable antibody fragment dimerized by the dHLX peptide with enhanced affinity against amantadine compared to its corresponding scFv antibody |
| title_full_unstemmed | A novel variable antibody fragment dimerized by the dHLX peptide with enhanced affinity against amantadine compared to its corresponding scFv antibody |
| title_short | A novel variable antibody fragment dimerized by the dHLX peptide with enhanced affinity against amantadine compared to its corresponding scFv antibody |
| title_sort | novel variable antibody fragment dimerized by the dhlx peptide with enhanced affinity against amantadine compared to its corresponding scfv antibody |
| topic | amantadine affinity binding ability scfv scfv-dhlx |
| url | http://dx.doi.org/10.1080/09540105.2017.1368459 |
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