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抗デオキシニバレノールモノクローナル抗体の作製と酵素サイクリング法を利用した高感度ELISAの確立
https://kagawa-u.repo.nii.ac.jp/records/3508
https://kagawa-u.repo.nii.ac.jp/records/3508c53c570f-1ea5-4e9b-a25b-f7d6af66c923
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
AN00038339_57_27.pdf (15.6 MB)
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| Item type | 紀要論文 / Departmental Bulletin Paper(1) | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 公開日 | 2012-03-27 | |||||||||||
| タイトル | ||||||||||||
| タイトル | 抗デオキシニバレノールモノクローナル抗体の作製と酵素サイクリング法を利用した高感度ELISAの確立 | |||||||||||
| 言語 | ja | |||||||||||
| タイトル | ||||||||||||
| タイトル | PRODUCTION MONOCLONAL ANTIBODIES AGAINST DEOXYNIVALENOL AND DEVELOPMENT OF A HIGH SENSITIVE ELISA USING ENZYME AMMPLIFICATION | |||||||||||
| 言語 | en | |||||||||||
| タイトル | ||||||||||||
| タイトル | コウ デオキシニバレノール モノクローナル コウタイ ノ サクセイ ト コウソ サイクリングホウ オ リヨウシタ コウカンド ELISA ノ カクリツ | |||||||||||
| 言語 | ja-Kana | |||||||||||
| 言語 | ||||||||||||
| 言語 | jpn | |||||||||||
| 資源タイプ | ||||||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||||||||
| 資源タイプ | departmental bulletin paper | |||||||||||
| アクセス権 | ||||||||||||
| アクセス権 | open access | |||||||||||
| アクセス権URI | http://purl.org/coar/access_right/c_abf2 | |||||||||||
| 著者 |
川村, 理
× 川村, 理
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| 抄録 | ||||||||||||
| 内容記述タイプ | Abstract | |||||||||||
| 内容記述 | In order to establish a high sensitive ELISA for deoxynivalenol (Don), worldwide occurring in cereal grains contaminated Fusarium graminearum, specific monoclonal antibodies (mAbs) against DON were newly prepared. The immunogen was prepared as follows ; 3-acetyl DON (3ADON) was reacted with anhydride hemiglutarated, and the resulted 3ADON hemigultarate (3ADON-HG) was deacylated by acetylase. The product, DON-HG, was coupled to bovine serum albumin (BSA) and ovalbumin by mixed anhydride reaction. Spleen cells from the mice immunized with DON-HG-BSA were fused with SP2/O myeloma. After HAT selection and cloning, five anti-DON antibody producing hybridomas (DON.1-5) were obtained. In the indirect competitive ELISA with horseradish peroxides-labeled second antibody and 3, 3', 5, 5' -tetramethly benzidine as substrate (HRP/TMBZ method), the detection limit was 100 ng/ml with DON. 1 or DON.3. The sensitive DON.1 and DON.3 cross-reacted with nivalenol (5 and 6%), and 15-acectyl DON (267 and 333%), respectively. DON.3 cross-reacted less than 0.5% other trichothecens. To improve the sensitivity, alkaline phosphatase-labeled second antibody and NADP as substrate (enzyme amplification method, ALP/NADP method) was established. The detection limit in the ALP/NDAP method using DON.3 was 5 ng/ml, 20-fold sensitive than the HRP/TMBZ method. Using the ALO/NDAP method, the quantification of DON in wheat and naked barley was performed. 70% aqueous methanol extract from wheat or naked barley was evaporated, and the residue dissolved in 0.01M phosphate buffered saline pH7.4 containing 0.05% Tween 20 and 10% methanol was subjected the ELISA. The detection limit was 50 ng/g. The average recovery of DON spiked in the range 50 to 2,500 ng/g was 105%. Survey natural occurrence has revealed that 18 out of 20 cereal samples in Japan were positive. The ELISA data were closely correlated with those gas chromatography (r^2=0.988). Application of the present ELISA in combination with an enzyme amplification system is expected for mass screenin | |||||||||||
| 言語 | en | |||||||||||
| 書誌情報 |
ja : 香川大学農学部学術報告 en : Technical bulletin of Faculty of Agriculture, Kagawa University 巻 57, p. 27-33, 発行日 2005-02-15 |
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| 出版者 | ||||||||||||
| 出版者 | 香川大学農学部 | |||||||||||
| 言語 | ja | |||||||||||
| ISSN | ||||||||||||
| 収録物識別子タイプ | PISSN | |||||||||||
| 収録物識別子 | 0368-5128 | |||||||||||
| 書誌レコードID | ||||||||||||
| 収録物識別子タイプ | NCID | |||||||||||
| 収録物識別子 | AN00038339 | |||||||||||
| 著者版フラグ | ||||||||||||
| 出版タイプ | VoR | |||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||||||||
| キーワード | ||||||||||||
| 言語 | en | |||||||||||
| 主題Scheme | Other | |||||||||||
| 主題 | Deoxynivalenol | |||||||||||
| キーワード | ||||||||||||
| 言語 | en | |||||||||||
| 主題Scheme | Other | |||||||||||
| 主題 | Monoclonal antibody | |||||||||||
| キーワード | ||||||||||||
| 言語 | en | |||||||||||
| 主題Scheme | Other | |||||||||||
| 主題 | ELISA | |||||||||||
| キーワード | ||||||||||||
| 言語 | en | |||||||||||
| 主題Scheme | Other | |||||||||||
| 主題 | An enzyme amplification system | |||||||||||
| WAID | ||||||||||||
| 1823 | ||||||||||||
| KEID | ||||||||||||
| 1326 | ||||||||||||
