Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom
Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/N...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Hebing LIU [verfasserIn] Yu QIN [verfasserIn] Weiwei XING [verfasserIn] Licai MA [verfasserIn] |
|---|
| Format: |
E-Artikel |
|---|---|
| Sprache: |
Chinesisch |
| Erschienen: |
2022 |
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| Schlagwörter: |
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| Übergeordnetes Werk: |
In: Shipin gongye ke-ji - The editorial department of Science and Technology of Food Industry, 2022, 43(2022), 5, Seite 294-301 |
|---|---|
| Übergeordnetes Werk: |
volume:43 ; year:2022 ; number:5 ; pages:294-301 |
| Links: |
|---|
| DOI / URN: |
10.13386/j.issn1002-0306.2021060279 |
|---|
| Katalog-ID: |
DOAJ025938576 |
|---|
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| 520 | |a Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. | ||
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10.13386/j.issn1002-0306.2021060279 doi (DE-627)DOAJ025938576 (DE-599)DOAJ9c8d6e72839f45ffbfb537947a6cbd87 DE-627 ger DE-627 rakwb chi TP368-456 Hebing LIU verfasserin aut Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. mushroom amanitin ic-elisa Food processing and manufacture Yu QIN verfasserin aut Weiwei XING verfasserin aut Licai MA verfasserin aut In Shipin gongye ke-ji The editorial department of Science and Technology of Food Industry, 2022 43(2022), 5, Seite 294-301 (DE-627)DOAJ000150428 10020306 nnns volume:43 year:2022 number:5 pages:294-301 https://doi.org/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/article/9c8d6e72839f45ffbfb537947a6cbd87 kostenfrei http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/toc/1002-0306 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA AR 43 2022 5 294-301 |
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10.13386/j.issn1002-0306.2021060279 doi (DE-627)DOAJ025938576 (DE-599)DOAJ9c8d6e72839f45ffbfb537947a6cbd87 DE-627 ger DE-627 rakwb chi TP368-456 Hebing LIU verfasserin aut Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. mushroom amanitin ic-elisa Food processing and manufacture Yu QIN verfasserin aut Weiwei XING verfasserin aut Licai MA verfasserin aut In Shipin gongye ke-ji The editorial department of Science and Technology of Food Industry, 2022 43(2022), 5, Seite 294-301 (DE-627)DOAJ000150428 10020306 nnns volume:43 year:2022 number:5 pages:294-301 https://doi.org/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/article/9c8d6e72839f45ffbfb537947a6cbd87 kostenfrei http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/toc/1002-0306 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA AR 43 2022 5 294-301 |
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10.13386/j.issn1002-0306.2021060279 doi (DE-627)DOAJ025938576 (DE-599)DOAJ9c8d6e72839f45ffbfb537947a6cbd87 DE-627 ger DE-627 rakwb chi TP368-456 Hebing LIU verfasserin aut Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. mushroom amanitin ic-elisa Food processing and manufacture Yu QIN verfasserin aut Weiwei XING verfasserin aut Licai MA verfasserin aut In Shipin gongye ke-ji The editorial department of Science and Technology of Food Industry, 2022 43(2022), 5, Seite 294-301 (DE-627)DOAJ000150428 10020306 nnns volume:43 year:2022 number:5 pages:294-301 https://doi.org/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/article/9c8d6e72839f45ffbfb537947a6cbd87 kostenfrei http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/toc/1002-0306 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA AR 43 2022 5 294-301 |
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10.13386/j.issn1002-0306.2021060279 doi (DE-627)DOAJ025938576 (DE-599)DOAJ9c8d6e72839f45ffbfb537947a6cbd87 DE-627 ger DE-627 rakwb chi TP368-456 Hebing LIU verfasserin aut Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. mushroom amanitin ic-elisa Food processing and manufacture Yu QIN verfasserin aut Weiwei XING verfasserin aut Licai MA verfasserin aut In Shipin gongye ke-ji The editorial department of Science and Technology of Food Industry, 2022 43(2022), 5, Seite 294-301 (DE-627)DOAJ000150428 10020306 nnns volume:43 year:2022 number:5 pages:294-301 https://doi.org/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/article/9c8d6e72839f45ffbfb537947a6cbd87 kostenfrei http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/toc/1002-0306 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA AR 43 2022 5 294-301 |
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10.13386/j.issn1002-0306.2021060279 doi (DE-627)DOAJ025938576 (DE-599)DOAJ9c8d6e72839f45ffbfb537947a6cbd87 DE-627 ger DE-627 rakwb chi TP368-456 Hebing LIU verfasserin aut Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. mushroom amanitin ic-elisa Food processing and manufacture Yu QIN verfasserin aut Weiwei XING verfasserin aut Licai MA verfasserin aut In Shipin gongye ke-ji The editorial department of Science and Technology of Food Industry, 2022 43(2022), 5, Seite 294-301 (DE-627)DOAJ000150428 10020306 nnns volume:43 year:2022 number:5 pages:294-301 https://doi.org/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/article/9c8d6e72839f45ffbfb537947a6cbd87 kostenfrei http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021060279 kostenfrei https://doaj.org/toc/1002-0306 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA AR 43 2022 5 294-301 |
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TP368-456 Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom mushroom amanitin ic-elisa |
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misc TP368-456 misc mushroom misc amanitin misc ic-elisa misc Food processing and manufacture |
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misc TP368-456 misc mushroom misc amanitin misc ic-elisa misc Food processing and manufacture |
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misc TP368-456 misc mushroom misc amanitin misc ic-elisa misc Food processing and manufacture |
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Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom |
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Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom |
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Hebing LIU |
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Hebing LIU Yu QIN Weiwei XING Licai MA |
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Elektronische Aufsätze |
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Hebing LIU |
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10.13386/j.issn1002-0306.2021060279 |
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verfasserin |
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establishment of indirect competitive elisa method for detecting amanitin in mushroom |
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TP368-456 |
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Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom |
| abstract |
Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. |
| abstractGer |
Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. |
| abstract_unstemmed |
Objective: This study aims to establish an indirect competitive enzyme-linked immunosorbent assay for the detection of amanitin (AMA) in mushroom. Methods: In this study, the hapten was obtained by introducing 6-aminocaproic acid into the carboxyl position of the α-amanotropin molecule through EDC/NHS, and further coupled with different carrier proteins to prepare immunogens and coating antigen. Afterwards, Balb/c mice were immunized with the immunogen to prepare monoclonal antibodies. Based on the obtained antibody, an indirect competitive ELISA method for the detection of amanitin in mushroom was established by optimizing the working concentrations of coating antigen and antibody, coating conditions, blocking conditions, working concentrations of enzyme labeled secondary antibody and incubation time. Finally, the sensitivity, recovery rate, intra batch and inter batch variation of the established method were evaluated. Results: The molecular weight of the synthesized hapten in this study was 1033.12, and the coupling ratio of the immunogen identified by MALDI-TOF was about 10.03. Based on hybridoma technology, the IC50 of the mouse monoclonal antibody 13H4 was 1.91 μg/L. Based on the obtained monoclonal antibody, the detection limit of the established ic-ELISA method for amanitin in mushroom was 0.88 μg/kg, the recovery rate was 85.66%~113.05%, the intra-assay coefficient of variation was 5.35%~9.54%, and the inter-assay coefficient of variation was less than 15%. Conclusion: The ic-ELISA method established in this study had high accuracy, precision, sensitivity, and stable performance. It provided a simple, reliable and rapid detection method for the analysis of the toxicogen of sudden mushroom poisoning. |
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Establishment of Indirect Competitive ELISA Method for Detecting Amanitin in Mushroom |
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https://doi.org/10.13386/j.issn1002-0306.2021060279 https://doaj.org/article/9c8d6e72839f45ffbfb537947a6cbd87 http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021060279 https://doaj.org/toc/1002-0306 |
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Yu QIN Weiwei XING Licai MA |
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