The monoclonal antibody specific for the 4‐hydroxy‐2‐nonenal histidine adduct

FEBS Letters

Miyake, Naoki¹  Uchida, Koji²  Osawa, Toshihiko²  Hiai, Hiroshi¹  Kawakishi, Shunro²  Hagiwara, Maiko²  Toyokuni, Shinya¹ 

[1] Department of Pathology, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606, Japan;Laboratory of Food and Biodynamics, Nagoya University School of Agriculture, Nagoya 464-01, Japan

关键词: Reactive oxygen species;    Lipid peroxidation;    4-Hydroxy-2-nonenal;    Histidine;    Monoclonal antibody;    ROS;    reactive oxygen species;    HNE;    4-hydroxy-2-nonenal;    KLH;    keyhole limpet hemocyanin;    mAb;    monoclonal antibody;    BSA;    bovine serum albumin;    ELISA;    enzyme-linked immunosorbent assay;    GAPDH;    glyceraldehyde-3-phosphate dehydrogenase;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    ECL;    enhanced chemiluminescence;    PBS;    phosphate-buffered saline;   

DOI  :  10.1016/0014-5793(95)00033-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Monoclonal antibodies directed against proteins modified with the major membrane lipid peroxidation product, 4-hydroxy-2-nonenal, have been established and characterized. The monoclonal antibodies specific for HNE-modified proteins were raised by immunizing mice with a HNE-keyhole limpet hemocyanin conjugate. The resulting five monoclonal antibodies (mAbs HNEJ-1–5) recognized HNE-modified bovine serum albumin (BSA), but not native BSA in Western blot studies. Of the five mAbs, HNEJ-2 exhibited the highest affinity for HNE-modified proteins and a much higher affinity for the HNE-histidine adduct than the HNE-lysine or HNE-cysteine adducts. mAb HNEJ-2 did not cross-react with proteins that had been treated with other aldehydes, such as 1-hexenal, 2-hexenal, 4-hydroxy-2-hexenal, 2-nonenal, formaldehyde, or glutaraldehyde. These results suggest that the major epitope recognized by mAb HNEJ-2 is the Michael addition-type HNE-histidine adduct.

【 授权许可】

Unknown