Immunochemical detection of carboxymethylated Apo B-100 in copper-oxidized LDL.
The objective of this study was to detect carboxymethyllysine (CML) moieties in biological materials using immunological methods. We prepared a polyclonal antibody specific to the carboxymethylated protein. The antibody recognizes the CML moiety in protein. Protein-bound carboxymethylcysteine (CMS) is scarcely recognized by the antibody. By use of the antibody, the formation of carboxymethylated apo B in copper-oxidized low density lipoprotein (ox-LDL) was immunochemically confirmed. To clarify the source compounds of carboxymethylation, various aldehyde-modified BSAs were prepared, and their reactivities with the antibody were investigated. Among the aldehyde-modified BSAs used, the glyoxal-BSA adduct showed a strong antigenicity. As far as we know, this is the first report of the detection of the CML moiety in ox-LDL using immunological methods.
Kato
Y
Y
Department of Applied Biological Sciences, Nagoya University, Japan.
Tokunaga
K
K
Osawa
T
T
D000906:Antibodies; D000918:Antibody Specificity; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000032:analysis / Q000737:chemistry*; D003300:Copper; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D008077:Lipoproteins, LDL / Q000032:analysis / Q000737:chemistry*; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D012710:Serum Albumin, Bovine; D017392:Thiobarbituric Acid Reactive Substances
D000906:Antibodies; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D008077:Lipoproteins, LDL; D017392:Thiobarbituric Acid Reactive Substances; C099252:oxidized low density lipoprotein; D012710:Serum Albumin, Bovine; C048496:N(6)-carboxymethyllysine; D003300:Copper; D008239:Lysine