The monoclonal antibody specific for the 4-hydroxy-2-nonenal histidine adduct.
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.
Toyokuni
S
S
Department of Pathology, Faculty of Medicine, Kyoto University, Japan.
Miyake
N
N
Hiai
H
H
Hagiwara
M
M
Kawakishi
S
S
Osawa
T
T
Uchida
K
K
D000447:Aldehydes / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D005260:Female; D006433:Hemocyanins / Q000276:immunology; D006639:Histidine / Q000276:immunology; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D011506:Proteins / Q000276:immunology; D014407:Tumor Cells, Cultured
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D011506:Proteins; D006639:Histidine; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; C027576:4-hydroxy-2-nonenal