Preparation of Monoclonal Antibodies Specifically Reacting with the Trichothecene Mycotoxins Nivalenol and 15-Acetylnivalenol via the Introduction of a Linker Molecule into Its C-15 Position
Kyoko Noda,
Yuki Hirakawa,
Tomomi Nishino,
Ritsuto Sekizuka,
Marin Kishimoto,
Tomohiro Furukawa,
Sakiko Sawane,
Ayu Matsunaga,
Naoki Kobayashi,
Kazutoshi Sugita,
Kenji Oonaka,
Hiroko Kawakami,
Yuji Otsuka,
Tetsuya Yamamoto,
Toshihiro Yamamoto,
Taku Yoshiya,
Maiko Watanabe,
Machiko Saka,
Keiko Momma,
Masayo Kushiro,
Shiro Miyake
Affiliations
Kyoko Noda
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Yuki Hirakawa
Department of Food and Nutrition, Kyoto Women’s University, Kyoto 605-8501, Japan
Tomomi Nishino
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Ritsuto Sekizuka
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Marin Kishimoto
Department of Nutrition and Food Science, Ochanomizu University, Bunkyo 112-8610, Japan
Tomohiro Furukawa
National Agriculture and Food Research Organization, Tsukuba 305-8642, Japan
Sakiko Sawane
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Ayu Matsunaga
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Naoki Kobayashi
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Kazutoshi Sugita
Department of Veterinary Medicine, Azabu University, Sagamihara 252-5201, Japan
Kenji Oonaka
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Hiroko Kawakami
Peptide Institute, Inc., Osaka 567-0085, Japan
Yuji Otsuka
Peptide Institute, Inc., Osaka 567-0085, Japan
Tetsuya Yamamoto
Peptide Institute, Inc., Osaka 567-0085, Japan
Toshihiro Yamamoto
Peptide Institute, Inc., Osaka 567-0085, Japan
Taku Yoshiya
Peptide Institute, Inc., Osaka 567-0085, Japan
Maiko Watanabe
National Institute of Health Sciences, Kawasaki 210-9501, Japan
Machiko Saka
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Keiko Momma
Department of Food and Nutrition, Kyoto Women’s University, Kyoto 605-8501, Japan
Masayo Kushiro
National Agriculture and Food Research Organization, Tsukuba 305-8642, Japan
Shiro Miyake
Department of Food and Life Science, Azabu University, Sagamihara 252-5201, Japan
Nivalenol (NIV) is a trichothecene mycotoxin that is more toxic than deoxynivalenol. It accumulates in grains due to infection with Fusarium species, which are the causative agents of scab or Fusarium head blight. An immunoassay, which is a rapid and easy analytical method, is necessary for monitoring NIV in grains. However, a specific antibody against NIV has not been prepared previously. To establish an immunoassay, we prepared NIV, introduced a linker, and generated antibodies against it. NIV was prepared from a culture of Fusarium kyushuense obtained from pressed barley through chromatographic procedures with synthetic adsorbents and silica gel. NIV was reacted with glutaric anhydride, and the reaction was stopped before mono-hemiglutaryl-NIV was changed to di-hemiglutaryl-NIV. 15-O-Hemiglutaryl-NIV was isolated via preparative HPLC and bound to keyhole limpet hemocyanin (KLH) using the active ester method. Two different monoclonal antibodies were prepared by immunizing mice with the NIV-KLH conjugate. The 50% inhibitory concentration values were 36 and 37 ng/mL. These antibodies also showed high reactivity in a direct competitive enzyme-linked immunosorbent assay and specifically reacted with NIV and 15-acetyl-NIV but not with deoxynivalenol and 4-acetyl-NIV.
