Monoclonal antibody based enzyme-linked immunosorbent assay for the specific detection of ciprofloxacin and enrofloxacin residues in fishery products
Ciprofloxacin (CPFX) residues in fishery products were dangerous for human health and still remain a big challenge in China. However, there are a few available assays to detect specifically CPFX residues in fishery products. This study aimed at generating CPFX-specific monoclonal antibodies (mAbs) a...
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| Veröffentlicht in: | Aquaculture 2010-12, Vol.310 (1), p.8-12 |
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| creator | Hu, Kun Huang, Xuanyun Jiang, Yousheng Fang, Wei Yang, Xianle |
| description | Ciprofloxacin (CPFX) residues in fishery products were dangerous for human health and still remain a big challenge in China. However, there are a few available assays to detect specifically CPFX residues in fishery products. This study aimed at generating CPFX-specific monoclonal antibodies (mAbs) and developing a competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) for the sensitive and specific detection of CPFX residues in fishery products. Firstly, the CPFX was conjugated with BSA and the conjugates were immunized to Balb/c mice. After characterizing anti-CPFX sera, we generated hybridoma cell lines and found that one hybridoma clone, 1C9, produced anti-CPFX mAb. Characterization of IC9 mAb revealed that it belonged to IgG2a, with an affinity of 3.75
×
10
10
mol/l for CPFX. Furthermore, we developed ic-ELISA using 150
ng/ml of CPFX-OVA for coating antigen and 0.22
μg of mAb. We found that this assay detected CPFX at 45.25
ng/ml and had 84.60% of cross-activity to enrofloxacin (ENR), but not to ofloxacin, levofloxacin, difloxacin, sarafloxacin, furacilin, chloramphenicol and nalachite green. Using this assay, we detected the CPFX in fishery products prepared with recovery rates from 80.57% to 94.61% and relative standard deviation (RSD) varying from 1.78% to 8.02%, similar to that by HPLC analysis. These data, together the protocol for the preparation of fishery products, indicate that the ic-ELISA is able to detect CPFX and ENR residues in fishery products sensitively and specifically within 2
h. Therefore, this assay can be used for monitoring CPFX and ENR in commercial fishery products, improving the safety of commercial meats in China. |
| doi_str_mv | 10.1016/j.aquaculture.2010.08.008 |
| format | Article |
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×
10
10
mol/l for CPFX. Furthermore, we developed ic-ELISA using 150
ng/ml of CPFX-OVA for coating antigen and 0.22
μg of mAb. We found that this assay detected CPFX at 45.25
ng/ml and had 84.60% of cross-activity to enrofloxacin (ENR), but not to ofloxacin, levofloxacin, difloxacin, sarafloxacin, furacilin, chloramphenicol and nalachite green. Using this assay, we detected the CPFX in fishery products prepared with recovery rates from 80.57% to 94.61% and relative standard deviation (RSD) varying from 1.78% to 8.02%, similar to that by HPLC analysis. These data, together the protocol for the preparation of fishery products, indicate that the ic-ELISA is able to detect CPFX and ENR residues in fishery products sensitively and specifically within 2
h. Therefore, this assay can be used for monitoring CPFX and ENR in commercial fishery products, improving the safety of commercial meats in China.</description><identifier>ISSN: 0044-8486</identifier><identifier>EISSN: 1873-5622</identifier><identifier>DOI: 10.1016/j.aquaculture.2010.08.008</identifier><identifier>CODEN: AQCLAL</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Animal aquaculture ; Animal productions ; Animal, plant and microbial ecology ; antibiotic residues ; Antigens ; Applied ecology ; Aquaculture ; Biological and medical sciences ; bovine serum albumin ; chloramphenicol ; Ciprofloxacin ; cross reaction ; detection ; drug residues ; enrofloxacin ; Enzyme-linked immunosorbent assay ; Exploitation and management of natural biological resources (hunting, fishing and exploited populations survey, etc.) ; Fish ; Fisheries ; Fishery products ; food safety ; Fundamental and applied biological sciences. Psychology ; General aspects ; hybridomas ; Immunoassay ; mice ; Monoclonal antibodies ; Monoclonal antibody ; ofloxacin ; rapid methods ; seafoods</subject><ispartof>Aquaculture, 2010-12, Vol.310 (1), p.8-12</ispartof><rights>2010</rights><rights>2015 INIST-CNRS</rights><rights>Copyright Elsevier Sequoia S.A. Dec 22, 2010</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c434t-c0b6979577ab238b10929e858a2cc4fc37bca165ece546e5632ed0d8565e8b9d3</citedby><cites>FETCH-LOGICAL-c434t-c0b6979577ab238b10929e858a2cc4fc37bca165ece546e5632ed0d8565e8b9d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.aquaculture.2010.08.008$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27922,27923,45993</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23652246$$DView record in Pascal Francis$$Hfree_for_read</backlink></links><search><creatorcontrib>Hu, Kun</creatorcontrib><creatorcontrib>Huang, Xuanyun</creatorcontrib><creatorcontrib>Jiang, Yousheng</creatorcontrib><creatorcontrib>Fang, Wei</creatorcontrib><creatorcontrib>Yang, Xianle</creatorcontrib><title>Monoclonal antibody based enzyme-linked immunosorbent assay for the specific detection of ciprofloxacin and enrofloxacin residues in fishery products</title><title>Aquaculture</title><description>Ciprofloxacin (CPFX) residues in fishery products were dangerous for human health and still remain a big challenge in China. However, there are a few available assays to detect specifically CPFX residues in fishery products. This study aimed at generating CPFX-specific monoclonal antibodies (mAbs) and developing a competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) for the sensitive and specific detection of CPFX residues in fishery products. Firstly, the CPFX was conjugated with BSA and the conjugates were immunized to Balb/c mice. After characterizing anti-CPFX sera, we generated hybridoma cell lines and found that one hybridoma clone, 1C9, produced anti-CPFX mAb. Characterization of IC9 mAb revealed that it belonged to IgG2a, with an affinity of 3.75
×
10
10
mol/l for CPFX. Furthermore, we developed ic-ELISA using 150
ng/ml of CPFX-OVA for coating antigen and 0.22
μg of mAb. We found that this assay detected CPFX at 45.25
ng/ml and had 84.60% of cross-activity to enrofloxacin (ENR), but not to ofloxacin, levofloxacin, difloxacin, sarafloxacin, furacilin, chloramphenicol and nalachite green. Using this assay, we detected the CPFX in fishery products prepared with recovery rates from 80.57% to 94.61% and relative standard deviation (RSD) varying from 1.78% to 8.02%, similar to that by HPLC analysis. These data, together the protocol for the preparation of fishery products, indicate that the ic-ELISA is able to detect CPFX and ENR residues in fishery products sensitively and specifically within 2
h. Therefore, this assay can be used for monitoring CPFX and ENR in commercial fishery products, improving the safety of commercial meats in China.</description><subject>Animal aquaculture</subject><subject>Animal productions</subject><subject>Animal, plant and microbial ecology</subject><subject>antibiotic residues</subject><subject>Antigens</subject><subject>Applied ecology</subject><subject>Aquaculture</subject><subject>Biological and medical sciences</subject><subject>bovine serum albumin</subject><subject>chloramphenicol</subject><subject>Ciprofloxacin</subject><subject>cross reaction</subject><subject>detection</subject><subject>drug residues</subject><subject>enrofloxacin</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Exploitation and management of natural biological resources (hunting, fishing and exploited populations survey, etc.)</subject><subject>Fish</subject><subject>Fisheries</subject><subject>Fishery products</subject><subject>food safety</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>General aspects</subject><subject>hybridomas</subject><subject>Immunoassay</subject><subject>mice</subject><subject>Monoclonal antibodies</subject><subject>Monoclonal antibody</subject><subject>ofloxacin</subject><subject>rapid methods</subject><subject>seafoods</subject><issn>0044-8486</issn><issn>1873-5622</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><recordid>eNqNkU2O1DAQhSMEEs3AGTBIiFUa20kcZ4law480iAXM2nLKZcZN2u6xE0S4B_elWj1CI1asXC599V6pXlW9EHwruFBv9lt7u1hYpnnJuJWc-lxvOdcPqo3QfVN3SsqH1Ybztq11q9Xj6kkpe865Up3YVL8_pZhgStFOzMY5jMmtbLQFHcP4az1gPYX4nX7hcFhiKimPGGdmS7Er8ymz-QZZOSIEH4A5nBHmkCJLnkE45uSn9NNCiCR-UrzXyFiCW7Awqn0oN5hXRgNugbk8rR55OxV8dvdeVNfvLr_uPtRXn99_3L29qqFt2rkGPqqhH7q-t6Ns9Cj4IAfUnbYSoPXQ9CNYoToE7FqFnWokOu50Ry09Dq65qF6fdcn4lnaZzSEUwGmyEdNSDJF9L3nLiXz5D7lPS6arEUQOvRZcEjScIciplIzeHHM42Lwawc0pLrM39-Iyp7gM14biotlXdwa2gJ18thFC-SsgG9VJ2Srinp85b5Ox3zIx119IqOFiEINqTsTuTCBd7kfAbAoEjIAuZErHuBT-Y58_79--4w</recordid><startdate>20101222</startdate><enddate>20101222</enddate><creator>Hu, Kun</creator><creator>Huang, Xuanyun</creator><creator>Jiang, Yousheng</creator><creator>Fang, Wei</creator><creator>Yang, Xianle</creator><general>Elsevier B.V</general><general>Amsterdam: Elsevier Science</general><general>Elsevier</general><general>Elsevier Sequoia S.A</general><scope>FBQ</scope><scope>IQODW</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QR</scope><scope>7ST</scope><scope>7TN</scope><scope>7U7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H94</scope><scope>H95</scope><scope>H98</scope><scope>H99</scope><scope>L.F</scope><scope>L.G</scope><scope>M7N</scope><scope>P64</scope><scope>SOI</scope></search><sort><creationdate>20101222</creationdate><title>Monoclonal antibody based enzyme-linked immunosorbent assay for the specific detection of ciprofloxacin and enrofloxacin residues in fishery products</title><author>Hu, Kun ; Huang, Xuanyun ; Jiang, Yousheng ; Fang, Wei ; Yang, Xianle</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c434t-c0b6979577ab238b10929e858a2cc4fc37bca165ece546e5632ed0d8565e8b9d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Animal aquaculture</topic><topic>Animal productions</topic><topic>Animal, plant and microbial ecology</topic><topic>antibiotic residues</topic><topic>Antigens</topic><topic>Applied ecology</topic><topic>Aquaculture</topic><topic>Biological and medical sciences</topic><topic>bovine serum albumin</topic><topic>chloramphenicol</topic><topic>Ciprofloxacin</topic><topic>cross reaction</topic><topic>detection</topic><topic>drug residues</topic><topic>enrofloxacin</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Exploitation and management of natural biological resources (hunting, fishing and exploited populations survey, etc.)</topic><topic>Fish</topic><topic>Fisheries</topic><topic>Fishery products</topic><topic>food safety</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>General aspects</topic><topic>hybridomas</topic><topic>Immunoassay</topic><topic>mice</topic><topic>Monoclonal antibodies</topic><topic>Monoclonal antibody</topic><topic>ofloxacin</topic><topic>rapid methods</topic><topic>seafoods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hu, Kun</creatorcontrib><creatorcontrib>Huang, Xuanyun</creatorcontrib><creatorcontrib>Jiang, Yousheng</creatorcontrib><creatorcontrib>Fang, Wei</creatorcontrib><creatorcontrib>Yang, Xianle</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Chemoreception Abstracts</collection><collection>Environment Abstracts</collection><collection>Oceanic Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Aquaculture Abstracts</collection><collection>ASFA: Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environment Abstracts</collection><jtitle>Aquaculture</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hu, Kun</au><au>Huang, Xuanyun</au><au>Jiang, Yousheng</au><au>Fang, Wei</au><au>Yang, Xianle</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Monoclonal antibody based enzyme-linked immunosorbent assay for the specific detection of ciprofloxacin and enrofloxacin residues in fishery products</atitle><jtitle>Aquaculture</jtitle><date>2010-12-22</date><risdate>2010</risdate><volume>310</volume><issue>1</issue><spage>8</spage><epage>12</epage><pages>8-12</pages><issn>0044-8486</issn><eissn>1873-5622</eissn><coden>AQCLAL</coden><abstract>Ciprofloxacin (CPFX) residues in fishery products were dangerous for human health and still remain a big challenge in China. However, there are a few available assays to detect specifically CPFX residues in fishery products. This study aimed at generating CPFX-specific monoclonal antibodies (mAbs) and developing a competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) for the sensitive and specific detection of CPFX residues in fishery products. Firstly, the CPFX was conjugated with BSA and the conjugates were immunized to Balb/c mice. After characterizing anti-CPFX sera, we generated hybridoma cell lines and found that one hybridoma clone, 1C9, produced anti-CPFX mAb. Characterization of IC9 mAb revealed that it belonged to IgG2a, with an affinity of 3.75
×
10
10
mol/l for CPFX. Furthermore, we developed ic-ELISA using 150
ng/ml of CPFX-OVA for coating antigen and 0.22
μg of mAb. We found that this assay detected CPFX at 45.25
ng/ml and had 84.60% of cross-activity to enrofloxacin (ENR), but not to ofloxacin, levofloxacin, difloxacin, sarafloxacin, furacilin, chloramphenicol and nalachite green. Using this assay, we detected the CPFX in fishery products prepared with recovery rates from 80.57% to 94.61% and relative standard deviation (RSD) varying from 1.78% to 8.02%, similar to that by HPLC analysis. These data, together the protocol for the preparation of fishery products, indicate that the ic-ELISA is able to detect CPFX and ENR residues in fishery products sensitively and specifically within 2
h. Therefore, this assay can be used for monitoring CPFX and ENR in commercial fishery products, improving the safety of commercial meats in China.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><doi>10.1016/j.aquaculture.2010.08.008</doi><tpages>5</tpages></addata></record> |
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| source | Elsevier ScienceDirect Journals Complete |
| subjects | Animal aquaculture Animal productions Animal, plant and microbial ecology antibiotic residues Antigens Applied ecology Aquaculture Biological and medical sciences bovine serum albumin chloramphenicol Ciprofloxacin cross reaction detection drug residues enrofloxacin Enzyme-linked immunosorbent assay Exploitation and management of natural biological resources (hunting, fishing and exploited populations survey, etc.) Fish Fisheries Fishery products food safety Fundamental and applied biological sciences. Psychology General aspects hybridomas Immunoassay mice Monoclonal antibodies Monoclonal antibody ofloxacin rapid methods seafoods |
| title | Monoclonal antibody based enzyme-linked immunosorbent assay for the specific detection of ciprofloxacin and enrofloxacin residues in fishery products |
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