Monoclonal and polyclonal antibodies against acetaldehyde-containing epitopes in acetaldehyde-protein adducts.

Abstract

Immunization of mice with acetaldehyde conjugated to human plasma proteins resulted in the production of polyclonal antibodies that reacted with erythrocyte protein-acetaldehyde conjugates, but not with control erythrocyte proteins. Such antibodies recognized erythrocyte protein-acetaldehyde conjugates prepared with 20-100 microM acetaldehyde, concentrations that exist in the blood of alcoholics. The antibodies also recognized acetaldehyde condensation products with synthetic poly-(L-lysine). Immunization with keyhole limpet hemocyanin-acetaldehyde conjugates resulted in antibodies against both plasma protein-acetaldehyde and erythrocyte protein-acetaldehyde conjugates, which did not cross-react with the respective unmodified carrier proteins. Immunization with human erythrocyte protein-acetaldehyde condensates led to the production of antibodies against both the protein moiety as well as the condensate. Monoclonal antibodies with affinities 50 times greater for the condensate than for the carrier protein were produced by hybridization of spleen cells from the immunized mice. Chronic alcohol administration to mice for 45-50 days led to the generation of antibodies that reacted against protein-acetaldehyde conjugates, suggesting that such adducts are formed in vivo and can act as neoantigens. Antibodies against acetaldehyde adducts should be of value in the identification of alcohol consumption and in the study of the biology of the adducts in relation to organ pathology.

Immunological Detection of Acetaldehyde-Protein Adducts in Ethanol-Treated Carrot Cells

“…The authors named above hypothesized a role for acetaldehyde adducts to proteins in the pathogenesis of alcohol-induced liver injuries. Israel et al (8) showed that antibodies can be obtained against epitopes containing the acetaldehyde residue in acetaldehyde-protein condensates and that the antibodies can recognize the adducts regardless of the nature of the protein binding the acetaldehyde moiety.…”

“…Acetaldehyde binds to proteins leading to the formation of both stable and unstable adducts (8); the latter, postulated to be Schiffs bases, can be stabilized in vitro ' Research supported by National Research Council of Italy, Special Project RAISA, Subproject 2,paper No. 264.…”

A Competitive Enzyme-Linked Immunosorbent Assay to Quantify Acetaldehyde-Protein Adducts That Accumulate in Dry Seeds during Aging

“…The modification of seed proteins would cause proteins to denature, which may cause seed aging. In some in vitro experiments, it has been demonstrated that acetaldehyde attacks Lys residues (Tuma et al, 1987) and causes the loss of the catalytic activity of some enzymes (Mauch, et al, 1986).…”

“…The direct ELISA was tested for the assay of APA in a model system in which ovalbumin was used as a carrier protein for the synthesis of AOAs at acetaldehyde concen- trations from O to 1 mM (Israel et al, 1986;Perata et al, 1992). It was possible by the direct method to distinguish AOAl from other AOAs produced at lower acetaldehyde concentrations (Fig.…”

“…The toxic effects of acetaldehyde may arise from its nonenzymatic binding with proteins (Gaines et al, 1977;Stevens et al, 1981;Donohue et al, 1983) and DNA (Restow and Obe, 1978;Fraenkel-Conrat and Singer, 1988). Israel et al (1986) found that the immunization of mice with acetaldehyde conjugated to human plasma proteins results in the production of a polyclonal antibody, which reacts with various APAs. The production of acetaldehyde in tissue culture is thought to cause cell damage (Perata et al, 1988;Perata and Alpi, 1991).…”