p‑Bromophenol-Enhanced Bienzymatic Chemiluminescence Competitive Immunoassay for Ultrasensitive Determination of Aflatoxin B1
Aflatoxin B1 (AFB1) contamination is one of the most critical global issues in food safety. The high carcinogenic nature necessitates rapid and specific methods for the determination of AFB1 in foodstuffs at ultratrace levels. Here, we report an enhanced bienzymatic chemiluminescence competitive imm...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2019-10, Vol.91 (20), p.13191-13197 |
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| creator | Li, Juan Zhao, Xu Chen, Li-Jian Qian, Hai-Long Wang, Wen-Long Yang, Cheng Yan, Xiu-Ping |
| description | Aflatoxin B1 (AFB1) contamination is one of the most critical global issues in food safety. The high carcinogenic nature necessitates rapid and specific methods for the determination of AFB1 in foodstuffs at ultratrace levels. Here, we report an enhanced bienzymatic chemiluminescence competitive immunoassay for ultrasensitive and high-throughput determination of AFB1. In this assay, protein G was first coated on the wells of a microplate for recognizing the Fc fragment of anti-AFB1 mAbs to reduce the antibody dosage and guarantee high immunological reaction efficiency. The target AFB1 competed with glucose oxidase labeled AFB1 for the limited anti-AFB1 mAbs in the wells of the microplate. p-Bromophenol was employed as an enhancer to obtain intense and long-lasting chemiluminescence. The utilization of an enhancer and bienzymatic catalysts effectively improved the detection sensitivity. The developed method offered a good linearity over 5 orders of magnitude, a detection limit of 5 pg L–1, and a relative standard deviation of 1.9% for AFB1. The application of the developed method to the analysis of grain samples gave quantitative recoveries from 94.0% to 97.0%. The developed method provides a universal platform for high-throughput, ultrasensitive, and high specific detection of pollutants or nutrients in foods. |
| doi_str_mv | 10.1021/acs.analchem.9b03579 |
| format | Article |
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The high carcinogenic nature necessitates rapid and specific methods for the determination of AFB1 in foodstuffs at ultratrace levels. Here, we report an enhanced bienzymatic chemiluminescence competitive immunoassay for ultrasensitive and high-throughput determination of AFB1. In this assay, protein G was first coated on the wells of a microplate for recognizing the Fc fragment of anti-AFB1 mAbs to reduce the antibody dosage and guarantee high immunological reaction efficiency. The target AFB1 competed with glucose oxidase labeled AFB1 for the limited anti-AFB1 mAbs in the wells of the microplate. p-Bromophenol was employed as an enhancer to obtain intense and long-lasting chemiluminescence. The utilization of an enhancer and bienzymatic catalysts effectively improved the detection sensitivity. The developed method offered a good linearity over 5 orders of magnitude, a detection limit of 5 pg L–1, and a relative standard deviation of 1.9% for AFB1. The application of the developed method to the analysis of grain samples gave quantitative recoveries from 94.0% to 97.0%. The developed method provides a universal platform for high-throughput, ultrasensitive, and high specific detection of pollutants or nutrients in foods.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.9b03579</identifier><language>eng</language><publisher>Washington: American Chemical Society</publisher><subject>Aflatoxin B1 ; Aflatoxins ; Antibodies ; Carcinogens ; Catalysts ; Chemiluminescence ; Chemistry ; Competition ; Contamination ; Food contamination ; Food safety ; Glucose oxidase ; Immunoassay ; Immunology ; Laboratory equipment ; Linearity ; Nutrients ; Organic chemistry ; Pollutants ; Pollution detection ; Protein G</subject><ispartof>Analytical chemistry (Washington), 2019-10, Vol.91 (20), p.13191-13197</ispartof><rights>Copyright American Chemical Society Oct 15, 2019</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><orcidid>0000-0001-8671-8766 ; 0000-0001-9953-7681</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.9b03579$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.9b03579$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,777,781,27057,27905,27906,56719,56769</link.rule.ids></links><search><creatorcontrib>Li, Juan</creatorcontrib><creatorcontrib>Zhao, Xu</creatorcontrib><creatorcontrib>Chen, Li-Jian</creatorcontrib><creatorcontrib>Qian, Hai-Long</creatorcontrib><creatorcontrib>Wang, Wen-Long</creatorcontrib><creatorcontrib>Yang, Cheng</creatorcontrib><creatorcontrib>Yan, Xiu-Ping</creatorcontrib><title>p‑Bromophenol-Enhanced Bienzymatic Chemiluminescence Competitive Immunoassay for Ultrasensitive Determination of Aflatoxin B1</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Aflatoxin B1 (AFB1) contamination is one of the most critical global issues in food safety. The high carcinogenic nature necessitates rapid and specific methods for the determination of AFB1 in foodstuffs at ultratrace levels. Here, we report an enhanced bienzymatic chemiluminescence competitive immunoassay for ultrasensitive and high-throughput determination of AFB1. In this assay, protein G was first coated on the wells of a microplate for recognizing the Fc fragment of anti-AFB1 mAbs to reduce the antibody dosage and guarantee high immunological reaction efficiency. The target AFB1 competed with glucose oxidase labeled AFB1 for the limited anti-AFB1 mAbs in the wells of the microplate. p-Bromophenol was employed as an enhancer to obtain intense and long-lasting chemiluminescence. The utilization of an enhancer and bienzymatic catalysts effectively improved the detection sensitivity. The developed method offered a good linearity over 5 orders of magnitude, a detection limit of 5 pg L–1, and a relative standard deviation of 1.9% for AFB1. The application of the developed method to the analysis of grain samples gave quantitative recoveries from 94.0% to 97.0%. The developed method provides a universal platform for high-throughput, ultrasensitive, and high specific detection of pollutants or nutrients in foods.</description><subject>Aflatoxin B1</subject><subject>Aflatoxins</subject><subject>Antibodies</subject><subject>Carcinogens</subject><subject>Catalysts</subject><subject>Chemiluminescence</subject><subject>Chemistry</subject><subject>Competition</subject><subject>Contamination</subject><subject>Food contamination</subject><subject>Food safety</subject><subject>Glucose oxidase</subject><subject>Immunoassay</subject><subject>Immunology</subject><subject>Laboratory equipment</subject><subject>Linearity</subject><subject>Nutrients</subject><subject>Organic chemistry</subject><subject>Pollutants</subject><subject>Pollution detection</subject><subject>Protein G</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><recordid>eNpFkE1OwzAQhS0EEqVwAxaWWKeM7eZv2YYClZDYlHU0TSZqqsQOsYMoG7gCV-QkuGoRq5HmvfctPsauBUwESHGLhZ2gxqbYUDtJ16DCOD1hIxFKCKIkkadsBAAqkDHAObuwdgsgBIhoxD67n6_veW9a021ImyZY6A3qgko-r0l_7Fp0dcEzD66boa012YJ8zDPTduRqV78RX7btoA1aiztemZ6_NK5HS9oe4jty1PupJxnNTcVnVYPOvNeaz8UlO6uwsXR1vGO2ul-sssfg6flhmc2eApQqccE6VhRhLAgpnFYJRtNqrdIIYlXKkEIFEiEswygVMVBSpjBFjNJQksBUqFSN2c0B2_XmdSDr8q0Zeq_M5lJ5ShILSHwLDi0v9L8gIN9bzvfPP8v50bL6BQyedso</recordid><startdate>20191015</startdate><enddate>20191015</enddate><creator>Li, Juan</creator><creator>Zhao, Xu</creator><creator>Chen, Li-Jian</creator><creator>Qian, Hai-Long</creator><creator>Wang, Wen-Long</creator><creator>Yang, Cheng</creator><creator>Yan, Xiu-Ping</creator><general>American Chemical Society</general><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><orcidid>https://orcid.org/0000-0001-8671-8766</orcidid><orcidid>https://orcid.org/0000-0001-9953-7681</orcidid></search><sort><creationdate>20191015</creationdate><title>p‑Bromophenol-Enhanced Bienzymatic Chemiluminescence Competitive Immunoassay for Ultrasensitive Determination of Aflatoxin B1</title><author>Li, Juan ; Zhao, Xu ; Chen, Li-Jian ; Qian, Hai-Long ; Wang, Wen-Long ; Yang, Cheng ; Yan, Xiu-Ping</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a238t-b73e6a71eae54f8a64fb396073d25e5302a05d569170e8d904aa6952e1a91393</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Aflatoxin B1</topic><topic>Aflatoxins</topic><topic>Antibodies</topic><topic>Carcinogens</topic><topic>Catalysts</topic><topic>Chemiluminescence</topic><topic>Chemistry</topic><topic>Competition</topic><topic>Contamination</topic><topic>Food contamination</topic><topic>Food safety</topic><topic>Glucose oxidase</topic><topic>Immunoassay</topic><topic>Immunology</topic><topic>Laboratory equipment</topic><topic>Linearity</topic><topic>Nutrients</topic><topic>Organic chemistry</topic><topic>Pollutants</topic><topic>Pollution detection</topic><topic>Protein G</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, Juan</creatorcontrib><creatorcontrib>Zhao, Xu</creatorcontrib><creatorcontrib>Chen, Li-Jian</creatorcontrib><creatorcontrib>Qian, Hai-Long</creatorcontrib><creatorcontrib>Wang, Wen-Long</creatorcontrib><creatorcontrib>Yang, Cheng</creatorcontrib><creatorcontrib>Yan, Xiu-Ping</creatorcontrib><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Li, Juan</au><au>Zhao, Xu</au><au>Chen, Li-Jian</au><au>Qian, Hai-Long</au><au>Wang, Wen-Long</au><au>Yang, Cheng</au><au>Yan, Xiu-Ping</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>p‑Bromophenol-Enhanced Bienzymatic Chemiluminescence Competitive Immunoassay for Ultrasensitive Determination of Aflatoxin B1</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2019-10-15</date><risdate>2019</risdate><volume>91</volume><issue>20</issue><spage>13191</spage><epage>13197</epage><pages>13191-13197</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>Aflatoxin B1 (AFB1) contamination is one of the most critical global issues in food safety. The high carcinogenic nature necessitates rapid and specific methods for the determination of AFB1 in foodstuffs at ultratrace levels. Here, we report an enhanced bienzymatic chemiluminescence competitive immunoassay for ultrasensitive and high-throughput determination of AFB1. In this assay, protein G was first coated on the wells of a microplate for recognizing the Fc fragment of anti-AFB1 mAbs to reduce the antibody dosage and guarantee high immunological reaction efficiency. The target AFB1 competed with glucose oxidase labeled AFB1 for the limited anti-AFB1 mAbs in the wells of the microplate. p-Bromophenol was employed as an enhancer to obtain intense and long-lasting chemiluminescence. The utilization of an enhancer and bienzymatic catalysts effectively improved the detection sensitivity. The developed method offered a good linearity over 5 orders of magnitude, a detection limit of 5 pg L–1, and a relative standard deviation of 1.9% for AFB1. The application of the developed method to the analysis of grain samples gave quantitative recoveries from 94.0% to 97.0%. The developed method provides a universal platform for high-throughput, ultrasensitive, and high specific detection of pollutants or nutrients in foods.</abstract><cop>Washington</cop><pub>American Chemical Society</pub><doi>10.1021/acs.analchem.9b03579</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0001-8671-8766</orcidid><orcidid>https://orcid.org/0000-0001-9953-7681</orcidid></addata></record> |
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| subjects | Aflatoxin B1 Aflatoxins Antibodies Carcinogens Catalysts Chemiluminescence Chemistry Competition Contamination Food contamination Food safety Glucose oxidase Immunoassay Immunology Laboratory equipment Linearity Nutrients Organic chemistry Pollutants Pollution detection Protein G |
| title | p‑Bromophenol-Enhanced Bienzymatic Chemiluminescence Competitive Immunoassay for Ultrasensitive Determination of Aflatoxin B1 |
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