Abstract
Unsaturated aldehydes such as acrolein and hydroxyalkenals are produced in vivo through lipid peroxidation chain reactions under conditions of oxidative stress or carcinogenic insult, and are causally involved in the pathogenesis of numerous diseases (1,2). The ability of cytotoxic and genotoxic compounds to inactivate various biologically important macromolecules has been well documented (3–5). For enzymes and proteins, lysine, cysteine, histidine, and other amino acid residues that play key roles in their functionings are preferentially modified. The complicated chemistry and biochemistry of the reactive molecules occurring in vivo in micro- or ultramicroquantities remain unclear, because direct determination of such unstable compounds is problematic. An alternative approach is the immunochemical quantification of reaction products accumulating in vivo, for which monoclonal antibodies (MAbs) specific for the acrolein-modified lysine and 4-hydroxy-2-nonenal (HNE)-modified histidine epitopes have been prepared (Fig. 1) (6,7). However, several difficulties are encountered with enzyme-linked immunosorbent assay (ELISA) determination of haptenic molecules. One is the low avidity of MAbs against the small epitopes in general. The other is that the most widely used sandwich method is unapplicable to the quantitation because monovalent antigens are undetectable in principle.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Hallwell, B. and Gutteridge, J. M. C. (1998) Free Radicals in Biology and Medicine, 3rd ed. Oxford Science Publ. Great Clarendon.
Esterbauer, H., Schaur, R. J., and Zollner, H. (1991) Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic. Biol. Med. 11, 81–128.
Benedetti, A., Comporti, M., and Esterbauer, H. (1980) Identification of 4-hydroxynonenal as a cytotoxic product originating from the peroxidation of liver microsomal lipids. Biochim. Biophys. Acta 620, 281–296.
Uchida, K. and Stadtman, E. R. (1992) Modification of histidine residues in proteins by reaction with 4-hydroxynonenal. Proc. Natl. Acad Sci. USA 89, 4544–4548.
Satoh, K., Kitahara, A., Soma, Y., Inaba, Y., Hatayama, I., and Sato, K. (1985) Purification, induction, and distribution of placental glutathione transferase: a new marker enzyme for preneoplastic cells in the rat chemical hepatocarcinogenesis. Proc. Natl. Acad. Sci. USA 82, 3964–3968.
Toyokuni, S., Miyake, N., Hiai, H., Hagiwara, M., Kawakishi, S., Osawa, T., and Uchida, K. (1995) The monoclonal antibody specific for the 4-hydroxy-2-nonenal histidine adduct. FEBS Lett. 359, 189–191.
Uchida, K., Kanematsu, M., Sakai, K., et al. (1998) Protein-bound acrolein: potential markers for oxidative stress. Proc. Natl. Acad. Sci. USA 95, 4852–4887.
Satoh, K., Yamada, S., Koike, Y., et al. (1999) A 1-hour enzyme-linked immunosorbent assay for quantitation of acrolein-and hydroxynonenal-modified proteins by epitope-bound casein matrix method. Anal. Biochem. 270, 323–328.
Uchida, K., Kanematsu, M., Morimitsu, Y., Osawa, T., Noguchi, N., and Niki, E. (1998) Acrolein is a product of lipid peroxidation reaction. Formation of free acrolein and its conjugate with lysine residues in oxidized low density lipoproteins. J. Biol. Chem. 273, 16,058–16,066.
Duhamel, R. C. and Johnson, D. A. (1985) Use of nonfat dry milk to block nonspecific nuclear and membrane staining by avidin conjugates. J. Histochem. Cytochem. 33, 711–714.
Froese, A., Sehon, A. H., and Eigen, M. (1962) Kinetic studies of protein-dye and antibody-hapten interactions with the temperature-jump method. Canad. J. Chem. 40, 1786–1797.
Day, L. A., Sturtevant, J. M., and Singer, S. J. (1962) The kinetics of the reactions between antibodies to the 2,4 dinitrophenyl group and specific haptens. Ann. N. Y. Acad. Sci. 103, 611–625.
Kabat, E. A. and Mayer, N. M. (1961) Experimental immunochemistry, 2nd ed., Charles C. Thomas Publ., Springfield, IL, pp. 22–96.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.,Totowa, NJ
About this protocol
Cite this protocol
Satoh, K., Uchida, K. (2003). Immunoassays for Lipid Peroxidation End Products. In: Hensley, K., Floyd, R.A. (eds) Methods in Biological Oxidative Stress. Methods in Pharmacology and Toxicology. Humana Press. https://doi.org/10.1385/1-59259-424-7:49
Download citation
DOI: https://doi.org/10.1385/1-59259-424-7:49
Publisher Name: Humana Press
Print ISBN: 978-0-89603-815-8
Online ISBN: 978-1-59259-424-5
eBook Packages: Springer Protocols