Heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine B in fraudulent food
Abstract Background The high quality of antibody (Ab) is critical for an immunoassay; usually, an Ab with low affinity is often regarded as a “bad” one in the immunoassay development. How to use a “bad” Ab to develop a highly sensitive immunoassay is still a huge challenge. Methods In this study, a...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2021-03-01
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| Series: | Chemical and Biological Technologies in Agriculture |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40538-021-00211-0 |
| _version_ | 1818651499218075648 |
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| author | Jin Wang Xing Shen Peng Zhong Zhaodong Li Qiushi Tang Xinan Huang Anatoly V. Zherdev Boris B. Dzantiev Sergei A. Eremin Zhili Xiao Hongtao Lei Xiangmei Li |
| author_facet | Jin Wang Xing Shen Peng Zhong Zhaodong Li Qiushi Tang Xinan Huang Anatoly V. Zherdev Boris B. Dzantiev Sergei A. Eremin Zhili Xiao Hongtao Lei Xiangmei Li |
| author_sort | Jin Wang |
| collection | DOAJ |
| description | Abstract Background The high quality of antibody (Ab) is critical for an immunoassay; usually, an Ab with low affinity is often regarded as a “bad” one in the immunoassay development. How to use a “bad” Ab to develop a highly sensitive immunoassay is still a huge challenge. Methods In this study, a heterologous immunoassay strategy was designed to enhance the sensitivity for the detection of banned dye, rhodamine B (RB), in fraudulent food. The RB Ab could not recognize RB by pairing with homologous coating antigen (Ag). However, the Ab showed unexpected high specificity and sensitivity recognition after being replaced by heterologous coating Ag. Indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed based on the heterologous strategy. Results The detection limit of icELISA for chilli powder, Chinese prickly ash, hot-pot seasoning, and chilli sauce was 0.002 μg/kg, and the recoveries of the four samples ranged from 76.0 to 102.0%, with the coefficient of variation between 3.9 and 18.8%. Parallel experiment for 20 market samples with high-performance liquid chromatography (HPLC) was performed on to confirm the performance of the practical application of the developed icELISA, and the results of the two methods had good correlation. Molecular modeling inferred that the carboxyl group of hapten and its exposure level played an important role in the hapten-Ab recognition. Conclusions The proposed icELISA can be used for the surveillance screening of RB in a range of seasoning foods, and the heterologous strategy is an effective approach to enhance the sensitivity in an immunoassay. |
| first_indexed | 2024-12-17T02:07:05Z |
| format | Article |
| id | doaj.art-e8ef4a717c824ea899648259f218de93 |
| institution | Directory Open Access Journal |
| issn | 2196-5641 |
| language | English |
| last_indexed | 2024-12-17T02:07:05Z |
| publishDate | 2021-03-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Chemical and Biological Technologies in Agriculture |
| spelling | doaj.art-e8ef4a717c824ea899648259f218de932022-12-21T22:07:40ZengSpringerOpenChemical and Biological Technologies in Agriculture2196-56412021-03-018111010.1186/s40538-021-00211-0Heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine B in fraudulent foodJin Wang0Xing Shen1Peng Zhong2Zhaodong Li3Qiushi Tang4Xinan Huang5Anatoly V. Zherdev6Boris B. Dzantiev7Sergei A. Eremin8Zhili Xiao9Hongtao Lei10Xiangmei Li11Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural UniversityGuangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural UniversityGuangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural UniversityGuangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural UniversityGuangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural UniversityTropical Medicine Institute & South China Chinese Medicine Collaborative Innovation Center, Guangzhou University of Chinese MedicineA.N. Bach Institute of Biochemistry, Federal Research Centre Fundamentals of Biotechnology, Russian Academy of SciencesA.N. Bach Institute of Biochemistry, Federal Research Centre Fundamentals of Biotechnology, Russian Academy of SciencesA.N. Bach Institute of Biochemistry, Federal Research Centre Fundamentals of Biotechnology, Russian Academy of SciencesGuangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural UniversityGuangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural UniversityGuangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural UniversityAbstract Background The high quality of antibody (Ab) is critical for an immunoassay; usually, an Ab with low affinity is often regarded as a “bad” one in the immunoassay development. How to use a “bad” Ab to develop a highly sensitive immunoassay is still a huge challenge. Methods In this study, a heterologous immunoassay strategy was designed to enhance the sensitivity for the detection of banned dye, rhodamine B (RB), in fraudulent food. The RB Ab could not recognize RB by pairing with homologous coating antigen (Ag). However, the Ab showed unexpected high specificity and sensitivity recognition after being replaced by heterologous coating Ag. Indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed based on the heterologous strategy. Results The detection limit of icELISA for chilli powder, Chinese prickly ash, hot-pot seasoning, and chilli sauce was 0.002 μg/kg, and the recoveries of the four samples ranged from 76.0 to 102.0%, with the coefficient of variation between 3.9 and 18.8%. Parallel experiment for 20 market samples with high-performance liquid chromatography (HPLC) was performed on to confirm the performance of the practical application of the developed icELISA, and the results of the two methods had good correlation. Molecular modeling inferred that the carboxyl group of hapten and its exposure level played an important role in the hapten-Ab recognition. Conclusions The proposed icELISA can be used for the surveillance screening of RB in a range of seasoning foods, and the heterologous strategy is an effective approach to enhance the sensitivity in an immunoassay.https://doi.org/10.1186/s40538-021-00211-0Rhodamine BELISAImmunoassayAntibodyHaptenHeterologous |
| spellingShingle | Jin Wang Xing Shen Peng Zhong Zhaodong Li Qiushi Tang Xinan Huang Anatoly V. Zherdev Boris B. Dzantiev Sergei A. Eremin Zhili Xiao Hongtao Lei Xiangmei Li Heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine B in fraudulent food Chemical and Biological Technologies in Agriculture Rhodamine B ELISA Immunoassay Antibody Hapten Heterologous |
| title | Heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine B in fraudulent food |
| title_full | Heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine B in fraudulent food |
| title_fullStr | Heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine B in fraudulent food |
| title_full_unstemmed | Heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine B in fraudulent food |
| title_short | Heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine B in fraudulent food |
| title_sort | heterologous immunoassay strategy for enhancing detection sensitivity of banned dye rhodamine b in fraudulent food |
| topic | Rhodamine B ELISA Immunoassay Antibody Hapten Heterologous |
| url | https://doi.org/10.1186/s40538-021-00211-0 |
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