The Effects of Different Antigen–Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in Vegetables
To establish rapid, high-sensitive, quantitative detection of ACP residues in vegetables. A 1G2 cell clone was selected as the most sensitive for anti-ACP antibody production following secondary immunization, cell fusion, and screening. The affinity of the 1G2 antibody to each of the four coating ag...
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MDPI AG
2022-09-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/12/9/730 |
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| author | Yuxiang Wu Yemin Guo Qingqing Yang Falan Li Xia Sun |
| author_facet | Yuxiang Wu Yemin Guo Qingqing Yang Falan Li Xia Sun |
| author_sort | Yuxiang Wu |
| collection | DOAJ |
| description | To establish rapid, high-sensitive, quantitative detection of ACP residues in vegetables. A 1G2 cell clone was selected as the most sensitive for anti-ACP antibody production following secondary immunization, cell fusion, and screening. The affinity of the 1G2 antibody to each of the four coating agents (imidacloprid–bovine serum albumin [BSA], thiacloprid–BSA, imidaclothiz–BSA, and ACP-BSA) was determined using a 20 min enzyme-linked immunosorbent assay (ELISA). The half maximal inhibitory concentration (IC<sub>50</sub>) was 0.51–0.62 ng/mL, showing no significant difference in affinity to different antigens. However, we obtained IC<sub>50</sub> values of 0.58 and 1.40 ng/mL on the linear regression lines for 1G2 anti-ACP antibody/imidacloprid–BSA and 1G2 anti-ACP antibody/thiacloprid–BSA, respectively, via quantum dot (QD)-based immunochromatography. That is, the 1G2 antibody/imidacloprid–BSA pair (the best combination) was about three times more sensitive than the 1G2 antibody/thiacloprid–BSA pair in immunochromatographic detection. The best combination was used for the development of an 8 min chromatographic paper test. With simple and convenient sample pretreatment, we achieved an average recovery of 75–117%. The coefficient of variation (CoV) was <25% for all concentrations tested, the false–positive rate was <5%, the false–negative rate was 0%, and the linear range of the method was 50–1800 μg/kg. These performance metrics met the ACP detection standards in China, the European Union (EU), and the United States (US). In summary, in this study, we established an 8 min QD-based immunochromatographic stripe for the rapid and accurate quantitative determination of ACP residues in vegetables. |
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| publishDate | 2022-09-01 |
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| series | Biosensors |
| spelling | doaj.art-f7e347e760904b1e8b43490131a9e18c2023-11-23T15:18:12ZengMDPI AGBiosensors2079-63742022-09-0112973010.3390/bios12090730The Effects of Different Antigen–Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in VegetablesYuxiang Wu0Yemin Guo1Qingqing Yang2Falan Li3Xia Sun4School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaSchool of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaSchool of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaSchool of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaSchool of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255049, ChinaTo establish rapid, high-sensitive, quantitative detection of ACP residues in vegetables. A 1G2 cell clone was selected as the most sensitive for anti-ACP antibody production following secondary immunization, cell fusion, and screening. The affinity of the 1G2 antibody to each of the four coating agents (imidacloprid–bovine serum albumin [BSA], thiacloprid–BSA, imidaclothiz–BSA, and ACP-BSA) was determined using a 20 min enzyme-linked immunosorbent assay (ELISA). The half maximal inhibitory concentration (IC<sub>50</sub>) was 0.51–0.62 ng/mL, showing no significant difference in affinity to different antigens. However, we obtained IC<sub>50</sub> values of 0.58 and 1.40 ng/mL on the linear regression lines for 1G2 anti-ACP antibody/imidacloprid–BSA and 1G2 anti-ACP antibody/thiacloprid–BSA, respectively, via quantum dot (QD)-based immunochromatography. That is, the 1G2 antibody/imidacloprid–BSA pair (the best combination) was about three times more sensitive than the 1G2 antibody/thiacloprid–BSA pair in immunochromatographic detection. The best combination was used for the development of an 8 min chromatographic paper test. With simple and convenient sample pretreatment, we achieved an average recovery of 75–117%. The coefficient of variation (CoV) was <25% for all concentrations tested, the false–positive rate was <5%, the false–negative rate was 0%, and the linear range of the method was 50–1800 μg/kg. These performance metrics met the ACP detection standards in China, the European Union (EU), and the United States (US). In summary, in this study, we established an 8 min QD-based immunochromatographic stripe for the rapid and accurate quantitative determination of ACP residues in vegetables.https://www.mdpi.com/2079-6374/12/9/730ACPvegetablesantigen–antibody pairsquantum dots8 min test |
| spellingShingle | Yuxiang Wu Yemin Guo Qingqing Yang Falan Li Xia Sun The Effects of Different Antigen–Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in Vegetables Biosensors ACP vegetables antigen–antibody pairs quantum dots 8 min test |
| title | The Effects of Different Antigen–Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in Vegetables |
| title_full | The Effects of Different Antigen–Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in Vegetables |
| title_fullStr | The Effects of Different Antigen–Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in Vegetables |
| title_full_unstemmed | The Effects of Different Antigen–Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in Vegetables |
| title_short | The Effects of Different Antigen–Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in Vegetables |
| title_sort | effects of different antigen antibody pairs on the results of 20 min elisa and 8 min chromatographic paper test for quantitative detection of acetamiprid in vegetables |
| topic | ACP vegetables antigen–antibody pairs quantum dots 8 min test |
| url | https://www.mdpi.com/2079-6374/12/9/730 |
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