IMMUNOREACTIVITY OF APOLIPOPROTEIN B-100 IN OXIDATIVELY MODIFIED LOW-DENSITY-LIPOPROTEIN

Thirteen monoclonal antibodies (MAbs) against apolipoprotein B-100 (ap e B) were used to analyze changes in immunoreactivity of human LDL res ulting from oxidation mediated by cupric ions and oxygen. Decrease in immunoreactivity of oxidized LDL was demonstrated by competitive ELISA with MAbs 5F8, BL3, Mb43, 2G8, B3, B5, and BL7 for which the epitopes are located within residues 1-1297, 4235-4355, 4027-4081, 3728-4306, 2239-2331, 1854-1878, and in the vicinity of residue 2331, respectivel y. Immunoreactivity of the epitope B6 (2239-2331) increased during fir st 4 hours of oxidation and then diminished gradually. Epitope B1 (405 -539) had slightly reduced immunoreactivity during first 8 h of LDL ox idation and then its minor increase was observed. MAb 12G10, specific to the epitope within ape B thrombin-digest fragment T4 (1-1297), disp layed either weak or strong binding to LDL. LDL with weak binding patt ern demonstrated significant increase in immunoreactivity upon oxidati on. In contrast, LDL with strong binding pattern showed little to no c hange. Epitopes Mb47 (3441-3569) and 8G4 (1-1297) remained unchanged i n oxidized LDL. Immunoreactivity of apo B-100 epitope recognized by MA b 4C11 (residues 2377-2658) was shown to be a function of oxidation ti me: it increased progressively up to 16 h and was stabilized for anoth er 24 h of LDL oxidation. This epitope may be unmasked by LDL oxidatio n and may provide a useful immunochemical marker to monitor the extent of LDL oxidation.