Monoclonal and polyclonal antibodies recognizing acetaldehyde-protein adducts.
Studies have investigated the hypothesis that metabolically derived acetaldehyde (AA) is capable of complexing with liver cell proteins to form AA-protein adducts that are capable of acting as antigens and inducing an immune response, as detected by the formation of unique antibodies. In an effort to better characterize and describe these adducts, mouse monoclonal and rabbit polyclonal antibodies specific for antigens prepared with AA under non-reducing (physiologic) and reducing (presence of sodium cyanoborohydride) conditions have been prepared. Two monoclonal antibodies were developed. The first antibody was RT1.1, which is specific to N-ethyl lysine (NEL); it is of the IgG2b isotype and recognizes all proteins modified with AA under reducing conditions. The other monoclonal antibody, NR-1, was of the IgG3 isotype; it recognizes proteins modified with AA under non-reducing conditions and cannot be inhibited by NEL. Affinity-purified and/or absorbed polyclonal antibodies were also produced to these epitopes. Using this panel of monoclonal and affinity-purified polyclonal antibodies, unique antigen-antibody binding occurred that: (1) detected only NEL; (2) reacted with the alpha-amino group on proteins prepared under reducing conditions; and (3) detected adducts on proteins prepared under non-reducing conditions. However, the only antibodies that recognized antigen(s) from alcohol-fed rat livers were those that were not specific to NEL or the alpha-amino group modified under reducing conditions. These data indicate that the relevant adduct in alcohol-fed rat livers is not NEL, and that it presumably is related to proteins modified with AA under non-reducing conditions.
Thiele
G M
GM
Veterans Administration Alcohol Research Center, Omaha Veterans Administration Medical Center, NE 68105, USA. lthiele@juno.com
Tuma
D J
DJ
Miller
J A
JA
Wegter
K M
KM
McDonald
T L
TL
Klassen
L W
LW
D000079:Acetaldehyde / Q000032:analysis / Q000378:metabolism*; D000437:Alcoholism / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis; D003600:Cytosol / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008239:Lysine; D051379:Mice; D008807:Mice, Inbred BALB C; D011506:Proteins / Q000276:immunology / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000941:Antigens; D011506:Proteins; D000079:Acetaldehyde; D008239:Lysine