Development of an Enzyme-Linked Immunosorbent Assay Based on a Monoclonal Antibody for the Rapid Detection of Citrinin in Wine
The ingestion of food contaminated with citrinin (CIT) poses a variety of health risks to humans and animals. The immunogens (CIT-COOH-BSA, CIT-H-BSA) and detection antigen (CIT-COOH-OVA, CIT-H-OVA) were synthesised using the active ester method (-COOH) and formaldehyde addition method (-H). A hybri...
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2024-12-01
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| author | Xingdong Yang Yang Qu Chenchen Wang Lihua Wu Xiaofei Hu |
| author_facet | Xingdong Yang Yang Qu Chenchen Wang Lihua Wu Xiaofei Hu |
| author_sort | Xingdong Yang |
| collection | DOAJ |
| description | The ingestion of food contaminated with citrinin (CIT) poses a variety of health risks to humans and animals. The immunogens (CIT-COOH-BSA, CIT-H-BSA) and detection antigen (CIT-COOH-OVA, CIT-H-OVA) were synthesised using the active ester method (-COOH) and formaldehyde addition method (-H). A hybridoma cell line (3G5) that secretes anti-CIT monoclonal antibodies (mAbs) was screened via CIT-H-BSA immunisation of mice, cell fusion, and ELISA screening technology. The cell line was injected intraperitoneally to prepare ascites. The reaction conditions for the indirect competitive ELISA (<i>ic</i>-ELISA) were optimised, and an <i>ic</i>-ELISA method for detecting CIT was preliminarily established. The results revealed that the IC<sub>50</sub> of CIT from optimised <i>ic</i>-ELISA was 37 pg/mL, the linear detection range was 5.9~230 pg/mL, and the cross-reaction (CR) rate with other analogues was less than 0.01%. The intra-assay and interassay sample recovery rates of CIT were 84.7~92.0% and 83.6~91.6%, and the coefficients of variation (CVs) were less than 10%. The <i>ic</i>-ELISA of CIT established in this study was not significantly different from the HPLC results and is rapid, highly sensitive and strongly specific, providing technical support for the detection of CIT. |
| format | Article |
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| institution | Kabale University |
| issn | 2304-8158 |
| language | English |
| publishDate | 2024-12-01 |
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| spelling | doaj-art-01b8391c2faf4dccacd17debdff2fff92025-01-10T13:17:33ZengMDPI AGFoods2304-81582024-12-011412710.3390/foods14010027Development of an Enzyme-Linked Immunosorbent Assay Based on a Monoclonal Antibody for the Rapid Detection of Citrinin in WineXingdong Yang0Yang Qu1Chenchen Wang2Lihua Wu3Xiaofei Hu4Institute of Food and Drug Inspection, College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou 466001, ChinaInstitute of Food and Drug Inspection, College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou 466001, ChinaInstitute of Food and Drug Inspection, College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou 466001, ChinaInstitute of Food and Drug Inspection, College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou 466001, ChinaKey Laboratory of Animal Immunology, Ministry of Agriculture and Rural Affairs & Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, ChinaThe ingestion of food contaminated with citrinin (CIT) poses a variety of health risks to humans and animals. The immunogens (CIT-COOH-BSA, CIT-H-BSA) and detection antigen (CIT-COOH-OVA, CIT-H-OVA) were synthesised using the active ester method (-COOH) and formaldehyde addition method (-H). A hybridoma cell line (3G5) that secretes anti-CIT monoclonal antibodies (mAbs) was screened via CIT-H-BSA immunisation of mice, cell fusion, and ELISA screening technology. The cell line was injected intraperitoneally to prepare ascites. The reaction conditions for the indirect competitive ELISA (<i>ic</i>-ELISA) were optimised, and an <i>ic</i>-ELISA method for detecting CIT was preliminarily established. The results revealed that the IC<sub>50</sub> of CIT from optimised <i>ic</i>-ELISA was 37 pg/mL, the linear detection range was 5.9~230 pg/mL, and the cross-reaction (CR) rate with other analogues was less than 0.01%. The intra-assay and interassay sample recovery rates of CIT were 84.7~92.0% and 83.6~91.6%, and the coefficients of variation (CVs) were less than 10%. The <i>ic</i>-ELISA of CIT established in this study was not significantly different from the HPLC results and is rapid, highly sensitive and strongly specific, providing technical support for the detection of CIT.https://www.mdpi.com/2304-8158/14/1/27citrininmonoclonal antibody<i>ic</i>-ELISAwine |
| spellingShingle | Xingdong Yang Yang Qu Chenchen Wang Lihua Wu Xiaofei Hu Development of an Enzyme-Linked Immunosorbent Assay Based on a Monoclonal Antibody for the Rapid Detection of Citrinin in Wine Foods citrinin monoclonal antibody <i>ic</i>-ELISA wine |
| title | Development of an Enzyme-Linked Immunosorbent Assay Based on a Monoclonal Antibody for the Rapid Detection of Citrinin in Wine |
| title_full | Development of an Enzyme-Linked Immunosorbent Assay Based on a Monoclonal Antibody for the Rapid Detection of Citrinin in Wine |
| title_fullStr | Development of an Enzyme-Linked Immunosorbent Assay Based on a Monoclonal Antibody for the Rapid Detection of Citrinin in Wine |
| title_full_unstemmed | Development of an Enzyme-Linked Immunosorbent Assay Based on a Monoclonal Antibody for the Rapid Detection of Citrinin in Wine |
| title_short | Development of an Enzyme-Linked Immunosorbent Assay Based on a Monoclonal Antibody for the Rapid Detection of Citrinin in Wine |
| title_sort | development of an enzyme linked immunosorbent assay based on a monoclonal antibody for the rapid detection of citrinin in wine |
| topic | citrinin monoclonal antibody <i>ic</i>-ELISA wine |
| url | https://www.mdpi.com/2304-8158/14/1/27 |
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