Sensitive Detection of Estrogenic Mycotoxin Zearalenone by Open Sandwich Immunoassay
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- SUZUKI Tatsuya
- Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo
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- MUNAKATA Yuriko
- Kyowa Medex Co.
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- MORITA Kazuki
- Kyowa Medex Co.
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- SHINODA Tatsuya
- Kyowa Medex Co.
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- UEDA Hiroshi
- Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo PRESTO, JST
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抄録
Zearalenone (ZEA) is an estrogenic mycotoxin produced by Fusarium sp., and its production on corn and small grains during storage has been of considerable concern. For sensitive ZEA detection, we applied an open sandwich (OS) immunoassay that can noncompetitively detect monovalent antigens utilizing an antigen-induced enhancement of the VH/VL interaction. We cloned the VH and VL cDNAs of anti-ZEA mAb to a split-Fv phagemid pKST2, and firstly both VH and VL fragments were displayed on M13 phage p9 and p7, respectively, using an amber suppressor, TG-1, as a host. The split-Fv phage showed specific binding to immobilized ZEA, which was well inhibited by free ZEA. Then, the VH/VL interaction and its antigen-dependency were analyzed using a non-suppressor HB2151 as a host to produce VH-displaying phage and his/myc-tagged soluble VL in the culture supernatant. By capturing VL with an anti-myc or -his antibody and probing bound VH-phage, ZEA was successfully detected with a superior detection limit as well as a wider working range than those of a competitive assay. Also, essentially the same results were reproduced with purified VH-alkaline phosphatase and MBP-VL fusion proteins.
収録刊行物
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- Analytical Sciences
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Analytical Sciences 23 (1), 65-70, 2007
社団法人 日本分析化学会
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詳細情報 詳細情報について
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- CRID
- 1390282679232043264
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- NII論文ID
- 130004441371
- 10018464132
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- NII書誌ID
- AA10500785
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- COI
- 1:STN:280:DC%2BD2s%2Fhs1Kisw%3D%3D
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- ISSN
- 13482246
- 09106340
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- NDL書誌ID
- 8598149
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- Web Site
- https://ndlsearch.ndl.go.jp/books/R000000004-I8598149
- https://link.springer.com/content/pdf/10.2116/analsci.23.65.pdf
- https://link.springer.com/article/10.2116/analsci.23.65/fulltext.html
- http://www.jstage.jst.go.jp/article/analsci/23/1/23_1_65/_pdf
- https://search.jamas.or.jp/link/ui/2007175374
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- 本文言語コード
- en
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- JaLC
- NDL
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